TY - THES A1 - Krampert, Laura T1 - Dynamics of cardiac neutrophil diversity in murine myocardial infarction T1 - Dynamik der Diversität kardialer neutrophiler Granulozyten im Mausmodell Herzinfarkt N2 - After myocardial infarction, an inflammatory response is induced characterized by a sterile inflammation, followed by a reparative phase in order to induce cardiac healing. Neutrophils are the first immune cells that enter the ischemic tissue. Neutrophils have various functions in the ischemic heart, such as phagocytosis, production of reactive oxygen species or release of granule components. These functions can not only directly damage cardiac tissue, but are also necessary for initiating reparative effects in post-ischemic healing, indicating a dual role of neutrophils in cardiac healing after infarction. In recent years, evidence has been growing that neutrophils show phenotypic and functional differences in distinct homeostatic and pathogenic settings. Preliminary data of my working group using single-cell RNA-sequencing revealed the time- dependent heterogeneity of neutrophils, with different populations showing distinct gene expression profiles in ischemic hearts of mice, including the time-dependent appearance of a SiglecFhigh neutrophil population. To better understand the dynamics of neutrophil heterogeneity in the ischemic heart, my work aimed to validate previous findings at the protein level, as well as to investigate whether the distinct neutrophil populations show functional differences. Furthermore, in vivo depletion experiments were performed in order to modulate circulating neutrophil levels. Hearts, blood, bone marrow and spleens were processed and analyzed from mice after 1 day and 3 days after the onset of cardiac ischemia and analyzed using flow cytometry. Results showed that the majority of cardiac neutrophils isolated at day 3 after myocardial infarction were SiglecFhigh, whereas nearly no SiglecFhigh neutrophils could be isolated from ischemic hearts at day 1 after myocardial infarction. No SiglecFhigh neutrophils could be found in the blood, spleen and bone marrow either after 1 day or 3 days after myocardial infarction, indicating that the SiglecFhigh state of neutrophils is unique to the ischemic cardiac tissue. When I compared SiglecFhigh and SiglecFlow neutrophils regarding their phagocytosis activity and ROS production, SiglecFhigh neutrophils showed a higher phagocytosis ability than their SiglecFlow counterparts, as well as higher ROS production capacity. In vivo depletion experiments could not achieve successful and efficient depletion of cardiac neutrophils either 1 day or 3 days after myocardial infarction, but led to a shift of a higher percentage of SiglecFhigh expressing neutrophils in the depletion group. Bone marrow neutrophil levels only showed partial depletion at day 3 after MI. Regarding blood neutrophils, depletion efficiently reduced circulating neutrophils at both time points, 1 and 3 days after MI. To summarize, this work showed the time-dependent presence of different neutrophil states in the ischemic heart. The main population of neutrophils isolated 3 days after MI showed a high expression of SiglecF, a unique state that could not be detected at different time points or other organs. These SiglecFhigh neutrophils showed functional differences regarding their phagocytosis ability and ROS production. Further investigation is needed to reveal what role these SiglecFhigh neutrophils could play within the ischemic heart. To better target neutrophil depletion in vivo, more efficient or different anti-neutrophil strategies are needed. N2 - Nach Myokardinfarkt kommt es zu einer Entzündungsantwort, die durch eine sterile Entzündung und nachfolgende reparative Phase gekennzeichnet ist, um eine kardiale Heilung zu initiieren. Neutrophile Granulozyten sind die ersten Immunzellen, die das ischämische Gewebe infiltrieren. Neutrophile Granulozyten haben verschiedene Funktionen im ischämischen Herz, wie beispielsweise Phagozytose, Produktion reaktiver Oxigenspezies oder Entleerung von Granulae. Diese Funktionen können nicht nur das kardiale Gewebe direkt schädigen, sondern sind auch notwendig, um die reparative Phase zu initiieren, was auf eine duale Rolle der neutrophilen Granulozyten hinsichtlich kardialer Heilung hinweist. Evidenz der letzten Jahre zeigt, dass neutrophile Granulzyten phänotypische und funktionelle Unterschiede zeigen, sowohl in Homöostase als auch in verschiedenen pathologischen Umgebungen. Vorangehende Ergebnisse meiner Arbeitsgruppe von Einzelzellsequenzierungen zeigten die zeitabhängige Heterogenität neutrophiler Granulozyten, die in verschiedenen Populationen und mit unterschiedlichen Expressions-Mustern in ischämischen Mäuseherzen auftauchten. Unter anderem zeigte sich das zeitabhängige Auftauchen einer SiglecFhigh NeutrophilenPopulation. Um die Dynamik der Neutrophilen-Heterogenität im ischämischen Herz besser zu verstehen, war es das Ziel meiner Arbeit, vorangehende Ergebnisse auf Proteinbasis zu bestätigen, und die verschiedenen Neutrophilen-Populationen hinsichtlich möglicher funktioneller Unterschiede zu untersuchen. Zusätzlich wurden in-vivo Depletionsversuche durchgeführt, um das Vorkommen zirkulierender neutrophiler Granulozyten zu modulieren und mögliche Änderungen hinsichtlich des Vorkommens neutrophiler Granulozyten im Herzen zu untersuchen. Herz, Blut, Knochenmark und Milz von Mäusen, die einen 1 oder 3 Tage alten Myokardinfarkt hatten, wurden prozessiert und mittels Durchflusszytometrie analysiert. Es zeigte sich, dass der überwiegende Anteil kardialer neutrophiler Granulozyten die an Tag 3 nach Myokardinfarkt isoliert wurden SiglecFhigh waren, wohingegen quasi keine SiglecFhigh neutrophile Granulozyten an Tag 1 nach Myokardinfarkt gefunden werden konnten. Es konnten keine SiglecFhigh neutrophile Granulozyten in Blut, Knochenmark oder Milz gefunden werden, weder 1 noch 3 Tage nach Myokardinfarkt, was darauf hinweist, dass der SiglecFhigh Status neutrophiler Granulozyten spezifisch für ischämisches Herzgewebe ist. Im Vergleich von SiglecFhigh und SiglecFlow neutrophilen Granulozyten hinsichtlich der Phagozytose und Produktion reaktiver Oxigenspezies, zeigten SiglecFhigh neutrophile Granulozyten sowohl eine höhere Phagozytose als auch eine höhere Produktion reaktiver Oxigenspezies. In vivo Depletionsversuche konnten keine komplette und effiziente Depletion kardialer neutrophiler Granulozyten sowohl an Tag 1 als auch an Tag 3 nach Myokardinfarkt erzielen, führten jedoch zu einem höheren Prozentsatz an SiglecFhigh neutrophilen Granulozyten in der Depletionsgruppe. Neutrophile Granulozyten im Knochenmark konnten nur an Tag 3 teilweise depletiert werden. Mit Blick auf neutrophile Granulozyten im Blut führte die Depletion zu einer effizienten Reduktion zirkulierender neutrophiler Granulozyten sowohl an Tag 1 als auch an Tag 3 nach Myokardinfarkt. Zusammenfassend zeigt diese Arbeit die zeitabhängige Präsenz verschiedener neutrophiler Granulozyten im ischämischen Herzen. Der größte Anteil neutrophiler Granulozyten, die an Tag 3 isoliert wurden, zeigten eine hohe Expression von SiglecF, einem spezifischen und einzigartigem Phänotypus, der weder zu anderen Zeitpunkten noch in anderen Organen gefunden wurde. Diese SiglecFhigh neutrophilen Granulozyten zeigten funktionelle Unterschiede hinsichtlich ihrer Phagozytose und Produktion reaktiver Oxigenspezies. Weitere Untersuchungen sind notwendig um aufzuzeigen, welche Rolle diese SiglecFhigh neutrophilen Granulozyten im ischämischen Herzen spielen könnten.Um eine effizientere Depletion neutrophiler Granulozyten in vivo zu erzielen, sind andere oder effizientere Anti-Neutrophilen Strategien notwendig. KW - Neutrophiler Granulozyt KW - neutrophils KW - Entzündung KW - Herzinfarkt KW - inflammation KW - myocardial infarction Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-349576 ER - TY - JOUR A1 - Dresen, Ellen A1 - Pimiento, Jose M. A1 - Patel, Jayshil J. A1 - Heyland, Daren K. A1 - Rice, Todd W. A1 - Stoppe, Christian T1 - Overview of oxidative stress and the role of micronutrients in critical illness JF - Journal of Parenteral and Enteral Nutrition N2 - Inflammation and oxidative stress represent physiological response mechanisms to different types of stimuli and injury during critical illness. Its proper regulation is fundamental to cellular and organismal survival and are paramount to outcomes and recovery from critical illness. A proper maintenance of the delicate balance between inflammation, oxidative stress, and immune response is crucial for resolution from critical illness with important implications for patient outcome. The extent of inflammation and oxidative stress under normal conditions is limited by the antioxidant defense system of the human body, whereas the antioxidant capacity is commonly significantly compromised, and serum levels of micronutrients and vitamins significantly depleted in patients who are critically ill. Hence, the provision of antioxidants and anti-inflammatory nutrients may help to reduce the extent of oxidative stress and therefore improve clinical outcomes in patients who are critically ill. As existing evidence of the beneficial effects of antioxidant supplementation in patients who are critically ill is still unclear, actual findings about the most promising anti-inflammatory and antioxidative candidates selenium, vitamin C, zinc, and vitamin D will be discussed in this narrative review. The existing evidence provided so far demonstrates that several factors need to be considered to determine the efficacy of an antioxidant supplementation strategy in patients who are critically ill and indicates the need for adequately designed multicenter prospective randomized control trials to evaluate the clinical significance of different types and doses of micronutrients and vitamins in selected groups of patients with different types of critical illness. KW - critical illness KW - vitamins KW - vitamin C KW - inflammation KW - medical nutrition therapy KW - oxidative stress KW - selenium KW - trace elements KW - micronutrients KW - vitamin D KW - zinc Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-318186 VL - 47 SP - S38 EP - S49 ER - TY - JOUR A1 - Glaser, Kirsten A1 - Kern, David A1 - Speer, Christian P. A1 - Schlegel, Nicolas A1 - Schwab, Michael A1 - Thome, Ulrich H. A1 - Härtel, Christoph A1 - Wright, Clyde J. T1 - Imbalanced inflammatory responses in preterm and term cord blood monocytes and expansion of the CD14\(^+\)CD16\(^+\) subset upon toll-like receptor stimulation JF - International Journal of Molecular Sciences N2 - Developmentally regulated features of innate immunity are thought to place preterm and term infants at risk of infection and inflammation-related morbidity. Underlying mechanisms are incompletely understood. Differences in monocyte function including toll-like receptor (TLR) expression and signaling have been discussed. Some studies point to generally impaired TLR signaling, others to differences in individual pathways. In the present study, we assessed mRNA and protein expression of pro- and anti-inflammatory cytokines in preterm and term cord blood (CB) monocytes compared with adult controls stimulated ex vivo with Pam3CSK4, zymosan, polyinosinic:polycytidylic acid, lipopolysaccharide, flagellin, and CpG oligonucleotide, which activate the TLR1/2, TLR2/6, TLR3, TLR4, TLR5, and TLR9 pathways, respectively. In parallel, frequencies of monocyte subsets, stimulus-driven TLR expression, and phosphorylation of TLR-associated signaling molecules were analyzed. Independent of stimulus, pro-inflammatory responses of term CB monocytes equaled adult controls. The same held true for preterm CB monocytes—except for lower IL-1β levels. In contrast, CB monocytes released lower amounts of anti-inflammatory IL-10 and IL-1ra, resulting in higher ratios of pro-inflammatory to anti-inflammatory cytokines. Phosphorylation of p65, p38, and ERK1/2 correlated with adult controls. However, stimulated CB samples stood out with higher frequencies of intermediate monocytes (CD14\(^+\)CD16\(^+\)). Both pro-inflammatory net effect and expansion of the intermediate subset were most pronounced upon stimulation with Pam3CSK4 (TLR1/2), zymosan (TR2/6), and lipopolysaccharide (TLR4). Our data demonstrate robust pro-inflammatory and yet attenuated anti-inflammatory responses in preterm and term CB monocytes, along with imbalanced cytokine ratios. Intermediate monocytes, a subset ascribed pro-inflammatory features, might participate in this inflammatory state. KW - neonatal immunology KW - inflammation KW - preterm infants KW - monocytes KW - cord blood KW - monocyte subsets KW - cytokines KW - Toll-like receptor signaling Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-311056 SN - 1422-0067 VL - 24 IS - 5 ER - TY - JOUR A1 - García-Fernández, Patricia A1 - Reinhold, Colette A1 - Üçeyler, Nurcan A1 - Sommer, Claudia T1 - Local inflammatory mediators involved in neuropathic pain JF - International Journal of Molecular Sciences N2 - Polyneuropathy (PNP) is a term to describe diseases of the peripheral nervous system, 50% of which present with neuropathic pain. In some types of PNP, pain is restricted to the skin distally in the leg, suggesting a local regulatory process leading to pain. In this study, we proposed a pro-inflammatory pathway mediated by NF-κB that might be involved in the development of pain in patients with painful PNP. To test this hypothesis, we have collected nerve and skin samples from patients with different etiologies and levels of pain. We performed RT-qPCR to analyze the gene expression of the proposed inflammatory pathway components in sural nerve and in distal and proximal skin samples. In sural nerve, we showed a correlation of TLR4 and TNFα to neuropathic pain, and an upregulation of TNFα in patients with severe pain. Patients with an inflammatory PNP also presented a lower expression of TRPV1 and SIRT1. In distal skin, we found a reduced expression of TLR4 and miR-146-5p, in comparison to proximal skin. Our findings thus support our hypothesis of local inflammatory processes involved in pain in PNP, and further show disturbed anti-inflammatory pathways involving TRPV1 and SIRT1 in inflammatory PNP. KW - polyneuropathy KW - pain KW - inflammation KW - NF-κB KW - TNFα Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-313613 SN - 1422-0067 VL - 24 IS - 9 ER - TY - JOUR A1 - Ouhaddi, Yassine A1 - Charbonnier, Baptiste A1 - Porge, Juliette A1 - Zhang, Yu-Ling A1 - Garcia, Isadora A1 - Gbureck, Uwe A1 - Grover, Liam A1 - Gilardino, Mirko A1 - Harvey, Edward A1 - Makhoul, Nicholas A1 - Barralet, Jake T1 - Development of neovasculature in axially vascularized calcium phosphate cement scaffolds JF - Journal of Functional Biomaterials N2 - Augmenting the vascular supply to generate new tissues, a crucial aspect in regenerative medicine, has been challenging. Recently, our group showed that calcium phosphate can induce the formation of a functional neo-angiosome without the need for microsurgical arterial anastomosis. This was a preclinical proof of concept for biomaterial-induced luminal sprouting of large-diameter vessels. In this study, we investigated if sprouting was a general response to surgical injury or placement of an inorganic construct around the vessel. Cylindrical biocement scaffolds of differing chemistries were placed around the femoral vein. A contrast agent was used to visualize vessel ingrowth into the scaffolds. Cell populations in the scaffold were mapped using immunohistochemistry. Calcium phosphate scaffolds induced 2.7–3 times greater volume of blood vessels than calcium sulphate or magnesium phosphate scaffolds. Macrophage and vSMC populations were identified that changed spatially and temporally within the scaffold during implantation. NLRP3 inflammasome activation peaked at weeks 2 and 4 and then declined; however, IL-1β expression was sustained over the course of the experiment. IL-8, a promoter of angiogenesis, was also detected, and together, these responses suggest a role of sterile inflammation. Unexpectedly, the effect was distinct from an injury response as a result of surgical placement and also was not simply a foreign body reaction as a result of placing a rigid bioceramic next to a vein, since, while the materials tested had similar microstructures, only the calcium phosphates tested elicited an angiogenic response. This finding then reveals a potential path towards a new strategy for creating better pro-regenerative biomaterials. KW - angiogenesis KW - axial vascularization KW - bioceramic KW - bioinorganic KW - calcium phosphate KW - NLRP3 KW - inflammation Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-304026 SN - 2079-4983 VL - 14 IS - 2 ER - TY - JOUR A1 - Schanbacher, Constanze A1 - Hermanns, Heike M. A1 - Lorenz, Kristina A1 - Wajant, Harald A1 - Lang, Isabell T1 - Complement 1q/tumor necrosis factor-related proteins (CTRPs): structure, receptors and signaling JF - Biomedicines N2 - Adiponectin and the other 15 members of the complement 1q (C1q)/tumor necrosis factor (TNF)-related protein (CTRP) family are secreted proteins composed of an N-terminal variable domain followed by a stalk region and a characteristic C-terminal trimerizing globular C1q (gC1q) domain originally identified in the subunits of the complement protein C1q. We performed a basic PubMed literature search for articles mentioning the various CTRPs or their receptors in the abstract or title. In this narrative review, we briefly summarize the biology of CTRPs and focus then on the structure, receptors and major signaling pathways of CTRPs. Analyses of CTRP knockout mice and CTRP transgenic mice gave overwhelming evidence for the relevance of the anti-inflammatory and insulin-sensitizing effects of CTRPs in autoimmune diseases, obesity, atherosclerosis and cardiac dysfunction. CTRPs form homo- and heterotypic trimers and oligomers which can have different activities. The receptors of some CTRPs are unknown and some receptors are redundantly targeted by several CTRPs. The way in which CTRPs activate their receptors to trigger downstream signaling pathways is largely unknown. CTRPs and their receptors are considered as promising therapeutic targets but their translational usage is still hampered by the limited knowledge of CTRP redundancy and CTRP signal transduction. KW - adiponectin KW - AMPK KW - C1q/TNF related protein (CTRP) KW - inflammation KW - metabolism Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-304136 SN - 2227-9059 VL - 11 IS - 2 ER - TY - THES A1 - Leinweber, Jonas T1 - Untersuchung zur pathophysiologischen Rolle und therapeutischen Relevanz der neuen Inhibitoren der plasmatischen Blutgerinnung Agaphelin und Ixolaris im experimentellen Schlaganfallmodell der Maus T1 - Characterization of the pathophysiological role and therapeutic relevance of the new inhibitors of plasmatic blood coagulation Agaphelin and Ixolaris in the model of ischemic stroke in mice N2 - Beim ischämischen Schlaganfall führt ein thrombotischer Verschluss von gehirnversorgenden Arterien zu einer akuten Durchblutungsstörung, mit der Folge von neurologischen Defiziten. Primäres Therapieziel ist es, diese Blutgerinnsel aufzulösen, um die Sauerstoffversorgung des Gehirns wiederherzustellen und den ischämischen Hirnschaden zu begrenzen. Dazu stehen die intravenösen Thrombolyse mit rt-PA (rekombinanter Gewebe-Plasminogen-Aktivator) sowie die endovaskuläre mechanische Thrombektomie zur Verfügung. Häufig kann ein Schlaganfall, trotz erfolgreicher Rekanalisation der Gefäße, zu einer weiteren Größenzunahme des Infarktes und neurologischen Defiziten bei den Patienten führen. Diese Größenzunahme beruht zum einen auf einem sich entwickelnden Hirnödem und zum anderen auf entzündlichen Prozessen. Zahlreiche Hinweise deuten darauf hin, dass der Schlaganfall ein Zusammenspiel aus thrombotischen und entzündlichen Ereignissen ist, ein Phänomen, das als Thromboinflammation bezeichnet wird. Aufgrund der begrenzten Behandlungsmöglichkeiten ist die Entwicklung neuer Therapieansätze für den ischämischen Schlaganfall besonders wichtig. Agaphelin und Ixolaris sind Proteine aus den Speicheldrüsen von Hämatophagen, für welche in früheren Studien eine starke antithrombotische Wirkung bei gleichzeitig geringem Blutungsrisiko nachgewiesen wurde. Diese möglichen antithrombotischen Effekte wurden in dieser Studie im Hinblick auf ihre Wirksamkeit und Sicherheit im Mausmodell der zerebralen Ischämie untersucht. Die Behandlung der Mäuse mit Agaphelin 1 Stunde nach transienter Okklusion der Arteria cerebri media (tMCAO) führte zu kleineren Schlaganfallvolumina und geringeren neurologischen Defiziten an Tag 1 nach dem Schlaganfall. Die Mortalität der Mäuse war bis Tag 7 deutlich gesunken. Aus klinischer Sicht ist ebenfalls relevant, dass der starke antithrombotische Effekt von Agaphelin im Mausmodell nicht mit einem erhöhten Risiko für intrazerebrale Blutungen einherging. Diesem protektiven Effekt von Agaphelin lagen eine verminderte intrazerebrale Thrombusbildung, eine abgeschwächte Entzündungsantwort und eine Stabilisierung der Blut-Hirn-Schranke sowie eine Reduzierung der Apoptose zugrunde. Nach der Gabe von Ixolaris 1 Stunde nach tMCAO waren zwar signifikant geringere Infarktgrößen messbar, diese führten allerdings nicht zu einer Verbesserung der neurologischen Defizite. Zudem verursachte die Gabe von Ixolaris schon 24 Stunden nach tMCAO erhebliche intrazerebrale Blutungen und auch die Mortalität der Mäuse war zu diesem Zeitpunkt bereits erhöht. Aufgrund dieser massiven Nebenwirkungen scheint Ixolaris kein geeigneter Kandidat für eine humane Anwendung zu sein. Bei Agaphelin hingegen könnte es sich um einen vielversprechenden Kandidaten für die Behandlung des ischämischen Schlaganfalls handeln. Vor einer möglichen Testung von Agaphelin in klinischen Studien, sind weitere translationale Untersuchungen notwendig, um ein noch präziseres Verständnis für die Wirksamkeit und Sicherheit von Agaphelin zu gewinnen. Insgesamt stellt die Hemmung thromboinflammatorischer Prozesse, ohne eine Erhöhung der Blutungskomplikationen, eine vielversprechende Option zur Behandlung des ischämischen Schlaganfalls dar. N2 - Thrombotic occlusion of cerebral vessels is an important process in pathogenesis of ischemic stroke resulting in lack of blood supply of the brain and neurological deficits. In order to restore the oxygenation of the brain and to limit brain injury, recanalization of the occluded vessels is the therapeutic main goal of stroke treatment. So far, the only proven pharmacological intervention for thrombolysis is the recombinant tissue-type plasminogen activator. For recanalization of larger arteries endovascular thrombectomy was established as a mechanic intervention. Nevertheless, despite successful recanalization ischemic brain damage and neurological deficits evolve. Increased size of infarct lesions develop due to brain edema and inflammatory processes. Moreover, there is evidence that inflammation and thrombosis are linked, which has led to the concept of thromboinflammation. Due to the limited treatment strategies in stroke management, the development of new therapeutic approaches for ischemic stroke is particularly important. Recent studies have shown that the hematophagous salivary gland proteins Agaphelin and Ixolaris exhibit multiple antithrombotic effects without promoting a risk of bleeding. To investigate the potentially safe antithrombotic effects, Agaphelin and Ixolaris were tested in a mouse model of transient middle cerebral artery occlusion (tMCAO). Treatment of mice with Agaphelin 1 hour after tMCAO resulted in smaller infarct volumes and an improved neurological function on day one after stroke. Up to seven days after stroke the mortality rate was significantly reduced. This protective effect was due to reduced local thrombus formation, a reduced inflammatory response and less severe blood-brain-barrier damage as well as reduced apoptosis. Moreover, it is important to mention, that the strong protective effect of Agaphelin was not linked to an increased risk of intracerebral bleeding. Treatment of mice with Ixolaris one hour after tMCAO leads to significantly smaller infarct sizes. However, neurologic deficits did not improve after treatment with Ixolaris. Furthermore, risk of intracerebral bleeding and mortality rates were significantly increased 24 hours after treatment with Ixolaris. Due to these severe side effects, Ixolaris does not seem to be an appropriate candidate for human therapy. Nevertheless, Agaphelin appears to be a promising component for ischemic stroke treatment, but further translational studies should be performed before testing Agaphelin in clinical stroke trials. Overall, the inhibition of thromboinflammatory effects without increased bleeding reflects a promising option for successful ischemic stroke treatment. KW - Schlaganfall KW - antithrombotic KW - inflammation KW - stroke KW - Agaphelin KW - Ixolaris KW - Thromboinflammation KW - Experimenteller Schlaganfall Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-252921 ER - TY - JOUR A1 - Magliocca, Giorgia A1 - Mone, Pasquale A1 - Di Iorio, Biagio Raffaele A1 - Heidland, August A1 - Marzocco, Stefania T1 - Short-chain fatty acids in Chronic Kidney Disease: focus on inflammation and oxidative stress regulation JF - International Journal of Molecular Sciences N2 - Chronic Kidney Disease (CKD) is a debilitating disease associated with several secondary complications that increase comorbidity and mortality. In patients with CKD, there is a significant qualitative and quantitative alteration in the gut microbiota, which, consequently, also leads to reduced production of beneficial bacterial metabolites, such as short-chain fatty acids. Evidence supports the beneficial effects of short-chain fatty acids in modulating inflammation and oxidative stress, which are implicated in CKD pathogenesis and progression. Therefore, this review will provide an overview of the current knowledge, based on pre-clinical and clinical evidence, on the effect of SCFAs on CKD-associated inflammation and oxidative stress. KW - chronic kidney disease KW - short-chain fatty acids KW - oxidative stress KW - inflammation KW - uremic toxins Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-284587 SN - 1422-0067 VL - 23 IS - 10 ER - TY - JOUR A1 - Hung, Sophia A1 - Dreher, Liane A1 - Diessner, Joachim A1 - Schwarz, Stefan A1 - Ohlsen, Knut A1 - Hertlein, Tobias T1 - MRSA infection in the thigh muscle leads to systemic disease, strong inflammation, and loss of human monocytes in humanized mice JF - Frontiers in Immunology N2 - MRSA (Methicillin-resistant Staphylococcus aureus) is the second-leading cause of deaths by antibiotic-resistant bacteria globally, with more than 100,000 attributable deaths annually. Despite the high urgency to develop a vaccine to control this pathogen, all clinical trials with pre-clinically effective candidates failed so far. The recent development of “humanized” mice might help to edge the pre-clinical evaluation closer to the clinical situation and thus close this gap. We infected humanized NSG mice (huNSG: (NOD)-scid IL2R\(_γ\)\(^{null}\) mice engrafted with human CD34+ hematopoietic stem cells) locally with S. aureus USA300 LAC* lux into the thigh muscle in order to investigate the human immune response to acute and chronic infection. These mice proved not only to be more susceptible to MRSA infection than wild-type or “murinized” mice, but displayed furthermore inferior survival and signs of systemic infection in an otherwise localized infection model. The rate of humanization correlated directly with the severity of disease and survival of the mice. Human and murine cytokine levels in blood and at the primary site of infection were strongly elevated in huNSG mice compared to all control groups. And importantly, differences in human and murine immune cell lineages surfaced during the infection, with human monocyte and B cell numbers in blood and bone marrow being significantly reduced at the later time point of infection. Murine monocytes in contrast behaved conversely by increasing cell numbers. This study demonstrates significant differences in the in vivo behavior of human and murine cells towards S. aureus infection, which might help to sharpen the translational potential of pre-clinical models for future therapeutic approaches. KW - humanized mice KW - MRSA - methicillin-resistant Staphylococcus aureus KW - monocyte KW - bacterial infection model KW - inflammation KW - NSG KW - staphylocccal infection/epidemiology Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-278050 SN - 1664-3224 VL - 13 ER - TY - JOUR A1 - Weider, Margareta A1 - Schlagenhauf, Ulrich A1 - Seefried, Lothar T1 - Oral health status of adult hypophosphatasia patients: A cross‐sectional study JF - Journal of Clinical Periodontology N2 - Aim This study evaluated the oral health status of adult patients with hypophosphatasia (HPP). Materials and Methods Parameters of oral health assessment comprised decayed/missing/filled teeth (DMFT) index, probing pocket depth and clinical attachment level (CAL) as well as documentation of tooth loss and periodontal health status according to CCD/AAP criteria. Findings were compared with national reference data (DMS V survey) reporting oral health status in age‐related controls. Within‐group comparisons were made between the HPP patients harbouring one versus two alkaline phosphatase liver/bone/kidney type (ALPL) gene variants. Results Of 80 HPP patients (64 female) with a mean age of 46.4 years (range 24–78) and one (n = 55) or two (n = 18) variants (n = 7 lacking testing) within the ALPL gene, those with two variants displayed substantially higher tooth loss rate (14.0 ± 9.3) than those affected by only one ALPL variant (4.1 ± 5.4), who did not differ substantially from healthy DMS V controls. While DMFT score and severe periodontal diseases (PDs) of HPP patients with one variant only increased with progressing age, the two‐variant sub‐cohort age independently exhibited increased DMFT scores and a higher rate of severe PDs. Conclusions HPP patients affected by two variants of the ALPL gene exhibited a higher risk of periodontitis and tooth loss than the general population, while patients with one variant developed clinically relevant oral disease symptoms with progressing ageing. KW - dental status KW - hypophosphatasia KW - inflammation KW - periodontal disease KW - tooth loss Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-293777 VL - 49 IS - 12 SP - 1253 EP - 1261 ER - TY - JOUR A1 - Spitzel, Marlene A1 - Wagner, Elise A1 - Breyer, Maximilian A1 - Henniger, Dorothea A1 - Bayin, Mehtap A1 - Hofmann, Lukas A1 - Mauceri, Daniela A1 - Sommer, Claudia A1 - Üçeyler, Nurcan T1 - Dysregulation of immune response mediators and pain-related ion channels is associated with pain-like behavior in the GLA KO mouse model of Fabry disease JF - Cells N2 - Fabry disease (FD) is a rare life-threatening disorder caused by deficiency of the alpha-galactosidase A (GLA) enzyme with a characteristic pain phenotype. Impaired GLA production or function leads to the accumulation of the cell membrane compound globotriaosylceramide (Gb3) in the neurons of the dorsal root ganglia (DRG) of FD patients. Applying immunohistochemistry (IHC) and quantitative real-time polymerase chain reaction (qRT PCR) analysis on DRG tissue of the GLA knockout (KO) mouse model of FD, we address the question of how Gb3 accumulation may contribute to FD pain and focus on the immune system and pain-associated ion channel gene expression. We show a higher Gb3 load in the DRG of young (<6 months) (p < 0.01) and old (≥12 months) (p < 0.001) GLA KO mice compared to old wildtype (WT) littermates, and an overall suppressed immune response in the DRG of old GLA KO mice, represented by a reduced number of CD206\(^+\) macrophages (p < 0.01) and lower gene expression levels of the inflammation-associated targets interleukin(IL)1b (p < 0.05), IL10 (p < 0.001), glial fibrillary acidic protein (GFAP) (p < 0.05), and leucine rich alpha-2-glycoprotein 1 (LRG1) (p < 0.01) in the DRG of old GLA KO mice compared to old WT. Dysregulation of immune-related genes may be linked to lower gene expression levels of the pain-associated ion channels calcium-activated potassium channel 3.1 (KCa3.1) and transient receptor potential ankyrin 1 channel (TRPA1). Ion channel expression might further be disturbed by impaired sphingolipid recruitment mediated via the lipid raft marker flotillin-1 (FLOT1). This impairment is represented by an increased number of FLOT1\(^+\) DRG neurons with a membranous expression pattern in old GLA KO mice compared to young GLA KO, young WT, and old WT mice (p < 0.001 each). Further, we provide evidence for aberrant behavior of GLA KO mice, which might be linked to dysregulated ion channel gene expression levels and disturbed FLOT1 distribution patterns. Behavioral testing revealed mechanical hypersensitivity in young (p < 0.01) and old (p < 0.001) GLA KO mice compared to WT, heat hypersensitivity in young GLA KO mice (p < 0.001) compared to WT, age-dependent heat hyposensitivity in old GLA KO mice (p < 0.001) compared to young GLA KO mice, and cold hyposensitivity in young (p < 0.001) and old (p < 0.001) GLA KO mice compared to WT, which well reflects the clinical phenotype observed in FD patients. KW - Fabry disease KW - globotriaosylceramide KW - inflammation KW - macrophages KW - cytokines KW - ion channels KW - flotillin-1 lipid rafts KW - pain-associated behavior KW - mouse model Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-275186 SN - 2073-4409 VL - 11 IS - 11 ER - TY - JOUR A1 - Schapovalova, Olesia A1 - Gorlova, Anna A1 - de Munter, Johannes A1 - Sheveleva, Elisaveta A1 - Eropkin, Mikhail A1 - Gorbunov, Nikita A1 - Sicker, Michail A1 - Umriukhin, Aleksei A1 - Lyubchyk, Sergiy A1 - Lesch, Klaus-Peter A1 - Strekalova, Tatyana A1 - Schroeter, Careen A. T1 - Immunomodulatory effects of new phytotherapy on human macrophages and TLR4- and TLR7/8-mediated viral-like inflammation in mice JF - Frontiers in Medicine N2 - Background While all efforts have been undertaken to propagate the vaccination and develop remedies against SARS-CoV-2, no satisfactory management of this infection is available yet. Moreover, poor availability of any preventive and treatment measures of SARS-CoV-2 in economically disadvantageous communities aggravates the course of the pandemic. Here, we studied a new immunomodulatory phytotherapy (IP), an extract of blackberry, chamomile, garlic, cloves, and elderberry as a potential low-cost solution for these problems given the reported efficacy of herbal medicine during the previous SARS virus outbreak. Methods The key feature of SARS-CoV-2 infection, excessive inflammation, was studied in in vitro and in vivo assays under the application of the IP. First, changes in tumor-necrosis factor (TNF) and lnteurleukin-1 beta (IL-1β) concentrations were measured in a culture of human macrophages following the lipopolysaccharide (LPS) challenge and treatment with IP or prednisolone. Second, chronically IP-pre-treated CD-1 mice received an agonist of Toll-like receptors (TLR)-7/8 resiquimod and were examined for lung and spleen expression of pro-inflammatory cytokines and blood formula. Finally, chronically IP-pre-treated mice challenged with LPS injection were studied for “sickness” behavior. Additionally, the IP was analyzed using high-potency-liquid chromatography (HPLC)-high-resolution-mass-spectrometry (HRMS). Results LPS-induced in vitro release of TNF and IL-1β was reduced by both treatments. The IP-treated mice displayed blunted over-expression of SAA-2, ACE-2, CXCL1, and CXCL10 and decreased changes in blood formula in response to an injection with resiquimod. The IP-treated mice injected with LPS showed normalized locomotion, anxiety, and exploration behaviors but not abnormal forced swimming. Isoquercitrin, choline, leucine, chlorogenic acid, and other constituents were identified by HPLC-HRMS and likely underlie the IP immunomodulatory effects. Conclusions Herbal IP-therapy decreases inflammation and, partly, “sickness behavior,” suggesting its potency to combat SARS-CoV-2 infection first of all via its preventive effects. KW - toll-like receptors KW - SARS-CoV-2 KW - inflammation KW - pro-inflammatory cytokines KW - mice Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-286301 SN - 2296-858X VL - 9 ER - TY - JOUR A1 - Karnati, Srikanth A1 - Guntas, Gulcan A1 - Rajendran, Ranjithkumar A1 - Shityakov, Sergey A1 - Höring, Marcus A1 - Liebisch, Gerhard A1 - Kosanovic, Djuro A1 - Ergün, Süleyman A1 - Nagai, Michiaki A1 - Förster, Carola Y. T1 - Quantitative lipidomic analysis of Takotsubo syndrome patients' serum JF - Frontiers in Cardiovascular Medicine N2 - Takotsubo syndrome (TTS), also known as the transient left ventricular apical ballooning syndrome, is in contemporary times known as novel acute cardiac syndrome. It is characterized by transient left ventricular apical akinesis and hyperkinesis of the basal left ventricular portions. Although the precise etiology of TTS is unknown, events like the sudden release of stress hormones, such as the catecholamines and the increased inflammatory status might be plausible causes leading to the cardiovascular pathologies. Recent studies have highlighted that an imbalance in lipid accumulation might promote a deviant immune response as observed in TTS. However, there is no information on comprehensive profiling of serum lipids of TTS patients. Therefore, we investigated a detailed quantitative lipid analysis of TTS patients using ES-MSI. Our results showed significant differences in the majority of lipid species composition in the TTS patients compared to the control group. Furthermore, the computational analyses presented was able to link the altered lipids to the pro-inflammatory cytokines and disseminate possible mechanistic pathways involving TNFα and IL-6. Taken together, our study provides an extensive quantitative lipidome of TTS patients, which may provide a valuable Pre-diagnostic tool. This would facilitate the elucidation of the underlying mechanisms of the disease and to prevent the development of TTS in the future. KW - TTS KW - inflammation KW - lipids KW - TNF-α KW - IL6 KW - PIK3R1 KW - NF-kappa-B KW - phosphatidylinositol Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-270832 SN - 2297-055X VL - 9 IS - 797154 ER - TY - JOUR A1 - Thal, Serge C. A1 - Smetak, Manuel A1 - Hayashi, Kentaro A1 - Förster, Carola Y. T1 - Hemorrhagic cerebral insults and secondary Takotsubo syndrome: findings in a novel in vitro model using human blood samples JF - International Journal of Molecular Sciences N2 - Intracranial hemorrhage results in devastating forms of cerebral damage. Frequently, these results also present with cardiac dysfunction ranging from ECG changes to Takotsubo syndrome (TTS). This suggests that intracranial bleeding due to subarachnoid hemorrhage (SAH) disrupts the neuro–cardiac axis leading to neurogenic stress cardiomyopathy (NSC) of different degrees. Following this notion, SAH and secondary TTS could be directly linked, thus contributing to poor outcomes. We set out to test if blood circulation is the driver of the brain–heart axis by investigating serum samples of TTS patients. We present a novel in vitro model combining SAH and secondary TTS to mimic the effects of blood or serum, respectively, on blood–brain barrier (BBB) integrity using in vitro monolayers of an established murine model. We consistently demonstrated decreased monolayer integrity and confirmed reduced Claudin-5 and Occludin levels by RT-qPCR and Western blot and morphological reorganization of actin filaments in endothelial cells. Both tight junction proteins show a time-dependent reduction. Our findings highlight a faster and more prominent disintegration of BBB in the presence of TTS and support the importance of the bloodstream as a causal link between intracerebral bleeding and cardiac dysfunction. This may represent potential targets for future therapeutic inventions in SAH and TTS. KW - Takotsubo syndrome KW - subarachnoid hemorrhage KW - inflammation KW - in vitro KW - blood KW - blood–brain barrier KW - human KW - patient KW - endothelial cells Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-288305 SN - 1422-0067 VL - 23 IS - 19 ER - TY - JOUR A1 - Higuchi, Takahiro A1 - Serfling, Sebastian E. A1 - Rowe, Steven P. A1 - Werner, Rudolf A. T1 - Therapeutic effects of lipid lowering medications on myocardial blood flow, inflammation, and sympathetic nerve activity using nuclear techniques JF - Current Cardiology Reports N2 - Purpose of Review Statins are routinely applied in patients with coronary artery disease, as they allow significantly to reduce blood cholesterol levels. Although those drugs are endorsed by current guidelines and prescribed routinely, a substantial portion of patients are still statin-intolerant and image-piloted strategies may then be helpful to identify patients that need further intensified treatment, e.g., to initiate treatment with proprotein convertase subtilisin / kexin type 9 inhibitors (PCSK9i). In addition, it has also been advocated that statins exhibit nonlipid, cardio-protective effects including improved cardiac nerve integrity, blood flow, and anti-inflammatory effects in congestive heart failure (HF) patients. Recent Findings In subjects after myocardial infarction treated with statins, \(^{123}\)I-metaiodobenzylguanidine (MIBG) scintigraphy has already revealed enhanced cardiac nerve function relative to patients without statins. In addition, all of those aforementioned statin-targeted pathways in HF can be visualized and monitored using dedicated cardiac radiotracers, e.g., \(^{123}\)I-MIBG or \(^{18}\)F-AF78 (for cardiac nerve function), \(^{18}\)F-flurpiridaz (to determine coronary flow) or \(^{68}\)Ga-PentixaFor (to detect inflammation). Summary Statins exhibit various cardio-beneficial effects, including improvement of cardiac nerve function, blood flow, and reduction of inflammation, which can all be imaged using dedicated nuclear cardiac radiotracers. This may allow for in vivo monitoring of statin-induced cardioprotection beyond lipid profiling in HF patients. KW - sympathetic nervous system KW - cardiac nerve KW - MIBG KW - inflammation KW - blood flow KW - statin Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-324599 VL - 24 IS - 12 ER - TY - JOUR A1 - Lauruschkat, Chris D. A1 - Etter, Sonja A1 - Schnack, Elisabeth A1 - Ebel, Frank A1 - Schäuble, Sascha A1 - Page, Lukas A1 - Rümens, Dana A1 - Dragan, Mariola A1 - Schlegel, Nicolas A1 - Panagiotou, Gianni A1 - Kniemeyer, Olaf A1 - Brakhage, Axel A. A1 - Einsele, Hermann A1 - Wurster, Sebastian A1 - Loeffler, Juergen T1 - Chronic occupational mold exposure drives expansion of Aspergillus-reactive type 1 and type 2 T-helper cell responses JF - Journal of Fungi N2 - Occupational mold exposure can lead to Aspergillus-associated allergic diseases including asthma and hypersensitivity pneumonitis. Elevated IL-17 levels or disbalanced T-helper (Th) cell expansion were previously linked to Aspergillus-associated allergic diseases, whereas alterations to the Th cell repertoire in healthy occupationally exposed subjects are scarcely studied. Therefore, we employed functional immunoassays to compare Th cell responses to A. fumigatus antigens in organic farmers, a cohort frequently exposed to environmental molds, and non-occupationally exposed controls. Organic farmers harbored significantly higher A. fumigatus-specific Th-cell frequencies than controls, with comparable expansion of Th1- and Th2-cell frequencies but only slightly elevated Th17-cell frequencies. Accordingly, Aspergillus antigen-induced Th1 and Th2 cytokine levels were strongly elevated, whereas induction of IL-17A was minimal. Additionally, increased levels of some innate immune cell-derived cytokines were found in samples from organic farmers. Antigen-induced cytokine release combined with Aspergillus-specific Th-cell frequencies resulted in high classification accuracy between organic farmers and controls. Aspf22, CatB, and CipC elicited the strongest differences in Th1 and Th2 responses between the two cohorts, suggesting these antigens as potential candidates for future bio-effect monitoring approaches. Overall, we found that occupationally exposed agricultural workers display a largely balanced co-expansion of Th1 and Th2 immunity with only minor changes in Th17 responses. KW - mold exposure KW - immunoassay KW - biomarker KW - Aspergillus KW - cytokines KW - inflammation KW - adaptive immunity KW - hypersensitivity Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-245202 SN - 2309-608X VL - 7 IS - 9 ER - TY - THES A1 - Frischholz, Sebastian T1 - Resveratrol Counteracts IL-1β-mediated Impairment of Extracellular Matrix Deposition in 3D Articular Chondrocyte Constructs T1 - Resveratrol wirkt der IL-1β-vermittelten Beeinträchtigung von Extrazellulärmatrix-Deposition in 3D Konstrukten aus artikulären Chondrozyten entgegen N2 - Articular cartilage is an exceptional connective tissue which by a network of fibrillar collagen and glycosaminoglycan (GAG) molecules allows both low- friction articulation and distribution of loads to the subchondral bone (Armiento et al., 2018, Ulrich-Vinther et al., 2003). Because of its very limited ability to self-repair, chondral defects following traumatic injury increase the risk for secondary osteoarthritis (OA) (Muthuri et al., 2011). Still, current OA treatments such as common nonsteroidal anti-inflammatory drugs (NSAIDs) and joint replacement primarily address end-stage symptoms (Tonge et al., 2014). As low-grade inflammation plays a pivotal role in the pathogenesis of OA (Robinson et al., 2016), there is a strong demand for novel therapeutic concepts, such as integrating application of anti-inflammatory agents into cartilage cell- based therapies in order to effectively treat OA affected joints in early disease stages. The polyphenolic phytoalexin resveratrol (RSV), found in the skin of red grapes, berries, and peanuts, has been shown to have effective anti-inflammatory properties (Shen et al., 2012). However, its long-term effects on 3D chondrocyte constructs cultured in an inflammatory environment with regard to tissue quality have remained unexplored so far. Therefore, in this study, pellets made from expanded porcine articular chondrocytes were cultured for 14 days with either the pro-inflammatory cytokine interleukin-1β (IL-1β) (1 - 10 ng/ml) or RSV (50 μM) alone, or a co-treatment with both agents. Constructs treated with chondrocyte medium only served as control. Treatment with IL-1β at 10 ng/ml resulted in a significantly smaller pellet size and reduced DNA content. However, RSV counteracted the IL-1β-induced decrease and significantly enhanced diameter and DNA content. Also, in terms of GAG deposition, treatment with IL-1β at 10 ng/ml resulted in a tremendous depletion of absolute GAG content and GAG/DNA. Again, RSV co-treatment counteracted the inflammatory stimulus and led to a partial recovery of GAG content. Histological analysis utilizing safranin-O staining confirmed these findings. Marked expression of the cartilage-degrading enzyme matrix metalloproteinase 13 (MMP13) was detected in IL-1β-treated pellets, but none upon RSV co- treatment. Moreover, co-treatment of IL-1β-challenged constructs with RSV significantly increased absolute collagen content. However, under non- inflammatory conditions, RSV induced gene expression and protein accumulation of collagen type X, a marker for undesirable hypertrophy. Taken together, in the present thesis, RSV was demonstrated to elicit marked beneficial effects on the extracellular matrix composition of 3D cartilaginous constructs in long-term inflammatory culture in vitro, but also induced hypertrophy under non-inflammatory conditions. Based on these findings, further experiments examining multiple concentrations of RSV under various inflammatory conditions appear desirable concerning potential therapeutic applicability in OA. N2 - Gelenkknorpel ermöglicht als spezielles Bindegewebe aus Kollagenfasern und Glykosaminoglykanen (GAG) sowohl die reibungsarme Beweglichkeit in Gelenken als auch die Lastübertragung auf angrenzende Knochen (Armiento et al., 2018, Ulrich-Vinther et al., 2003). Aufgrund der sehr begrenzten Fähigkeit zur intrinsischen Erneuerung erhöhen chondrale Defekte nach traumatischen Verletzungen das Risiko für sekundäre Arthrose (Osteoarthritis; OA) (Muthuri et al., 2011). Dennoch konzentrieren sich derzeitige Behandlungsansätze, einschließlich nichtsteroidaler Antirheumatika (NSAR) und des operativen Gelenkersatzes, hauptsächlich auf Symptome im Endstadium der Erkrankung (Tonge et al., 2014). Da eine geringgradige Entzündung eine entscheidende Rolle in der Pathogenese der Arthrose spielt (Robinson et al., 2016), besteht ein starker Bedarf an neuartigen Therapiekonzepten, wie der Kombination von anti- inflammatorischen Wirkstoffen mit knorpelzellbasierten Therapien, um von Arthrose betroffene Gelenke in frühen Krankheitsstadien wirksam zu behandeln. Das polyphenolische Phytoalexin Resveratrol (RSV), welches in der Schale roter Weintrauben, in Beeren und Erdnüssen vorkommt, besitzt starke entzündungshemmende Eigenschaften (Shen et al., 2012). Langzeiteffekte auf 3D-Knorpelkonstrukte unter inflammatorischen Bedingungen sind hinsichtlich der Gewebequalität jedoch bislang unerforscht geblieben. Daher wurden in der vorliegenden Studie Pellets aus expandierten porcinen Gelenkknorpelzellen über einen Zeitraum von 14 Tagen entweder mit dem pro-inflammatorischen Zytokin Interleukin-1β (IL-1β) (1 - 10 ng/ml) oder RSV (50 μM) allein, oder mit beiden Agenzien kombiniert behandelt. Konstrukte, welche nur serumfreies Chondrozytenmedium erhielten, dienten als Kontrolle. Die Behandlung mit IL- 1β in einer Konzentration von 10 ng/ml führte zu einem signifikant geringeren Durchmesser der Pellets sowie einem verringerten DNA-Gehalt. RSV wirkte dieser IL-1β-vermittelten Reduktion entgegen und steigerte signifikant sowohl Durchmesser als auch DNA-Gehalt der untersuchten Konstrukte. Auch in Bezug auf die Deposition von GAG-Molekülen führte die Kultur mit IL-1β (10 ng/ml) zu einer massiven Abnahme des absoluten GAG-Gehaltes und der GAG/DNA- Ratio. Abermals wirkte die gleichzeitige Behandlung mit RSV dem Entzündungsreiz deutlich entgegen und resultierte in einer partiellen Wiederherstellung des GAG-Gehaltes. Die histologische Analyse unter Verwendung von Safranin-O-Färbungen bestätigte diese Ergebnisse. Darüber hinaus manifestierte sich eine ausgeprägte Expression des knorpelabbauenden Enzyms Matrix-Metalloproteinase 13 (MMP13) in IL-1β behandelten Pellets, nicht jedoch in denen, die simultan mit RSV behandelt wurden. Außerdem resultierte die gleichzeitige Behandlung von IL-1β-stimulierten Konstrukten mit RSV in einer signifikanten Erhöhung des absoluten Kollagengehaltes. Unter nicht-inflammatorischen Bedingungen induzierte RSV die Genexpression und Proteinakkumulation von Kollagen Typ X, einem Marker für unerwünschte Hypertrophie. Zusammengefasst wurde in der vorliegenden Arbeit gezeigt, dass RSV deutliche positive Effekte auf die Extrazellulärmatrix von 3D- Knorpelkonstrukten in einer Langzeit-Entzündungskultur in vitro hervorruft, allerdings unter nicht-inflammatorischen Bedingungen Hypertrophie induziert. Basierend auf diesen Befunden erscheinen weitere Experimente zur Untersuchung unterschiedlicher RSV-Konzentrationen unter verschiedenen Entzündungsbedingungen hinsichtlich einer möglichen therapeutischen Anwendbarkeit bei OA wünschenswert. KW - Resveratrol KW - Interleukin 1-beta KW - Gelenkknorpel KW - Extrazelluläre Matrix KW - Osteoarthritis KW - IL-1β KW - articular chondrocytes KW - cartilage KW - cell-based therapy KW - extracellular matrix KW - inflammation KW - osteoarthritis KW - resveratrol Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-237453 ER - TY - JOUR A1 - Freitag‐Wolf, Sandra A1 - Munz, Matthias A1 - Junge, Olaf A1 - Graetz, Christian A1 - Jockel‐Schneider, Yvonne A1 - Staufenbiel, Ingmar A1 - Bruckmann, Corinna A1 - Lieb, Wolfgang A1 - Franke, Andre A1 - Loos, Bruno G. A1 - Jepsen, Søren A1 - Dommisch, Henrik A1 - Schaefer, Arne S. T1 - Sex‐specific genetic factors affect the risk of early‐onset periodontitis in Europeans JF - Journal of Clinical Periodontology N2 - Aims Various studies have reported that young European women are more likely to develop early‐onset periodontitis compared to men. A potential explanation for the observed variations in sex and age of disease onset is the natural genetic variation within the autosomal genomes. We hypothesized that genotype‐by‐sex (G × S) interactions contribute to the increased prevalence and severity. Materials and methods Using the case‐only design, we tested for differences in genetic effects between men and women in 896 North‐West European early‐onset cases, using imputed genotypes from the OmniExpress genotyping array. Population‐representative 6823 controls were used to verify that the interacting variables G and S were uncorrelated in the general population. Results In total, 20 loci indicated G × S associations (P < 0.0005), 3 of which were previously suggested as risk genes for periodontitis (ABLIM2, CDH13, and NELL1). We also found independent G × S interactions of the related gene paralogs MACROD1/FLRT1 (chr11) and MACROD2/FLRT3 (chr20). G × S‐associated SNPs at CPEB4, CDH13, MACROD1, and MECOM were genome‐wide‐associated with heel bone mineral density (CPEB4, MECOM), waist‐to‐hip ratio (CPEB4, MACROD1), and blood pressure (CPEB4, CDH13). Conclusions Our results indicate that natural genetic variation affects the different heritability of periodontitis among sexes and suggest genes that contribute to inter‐sex phenotypic variation in early‐onset periodontitis. KW - alveolar bone loss KW - gene × sex interaction KW - genetic risk KW - heritability KW - inflammation Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-262445 VL - 48 IS - 11 SP - 1404 EP - 1413 ER - TY - JOUR A1 - Grassinger, Julia Maria A1 - Floren, Andreas A1 - Müller, Tobias A1 - Cerezo-Echevarria, Argiñe A1 - Beitzinger, Christoph A1 - Conrad, David A1 - Törner, Katrin A1 - Staudacher, Marlies A1 - Aupperle-Lellbach, Heike T1 - Digital lesions in dogs: a statistical breed analysis of 2912 cases JF - Veterinary Sciences N2 - Breed predispositions to canine digital neoplasms are well known. However, there is currently no statistical analysis identifying the least affected breeds. To this end, 2912 canine amputated digits submitted from 2014–2019 to the Laboklin GmbH & Co. KG for routine diagnostics were statistically analyzed. The study population consisted of 155 different breeds (most common: 634 Mongrels, 411 Schnauzers, 197 Labrador Retrievers, 93 Golden Retrievers). Non-neoplastic processes were present in 1246 (43%), tumor-like lesions in 138 (5%), and neoplasms in 1528 cases (52%). Benign tumors (n = 335) were characterized by 217 subungual keratoacanthomas, 36 histiocytomas, 35 plasmacytomas, 16 papillomas, 12 melanocytomas, 9 sebaceous gland tumors, 6 lipomas, and 4 bone tumors. Malignant neoplasms (n = 1193) included 758 squamous cell carcinomas (SCC), 196 malignant melanomas (MM), 76 soft tissue sarcomas, 52 mast cell tumors, 37 non-specified sarcomas, 29 anaplastic neoplasms, 24 carcinomas, 20 bone tumors, and 1 histiocytic sarcoma. Predisposed breeds for SCC included the Schnauzer (log OR = 2.61), Briard (log OR = 1.78), Rottweiler (log OR = 1.54), Poodle (log OR = 1.40), and Dachshund (log OR = 1.30). Jack Russell Terriers (log OR = −2.95) were significantly less affected by SCC than Mongrels. Acral MM were significantly more frequent in Rottweilers (log OR = 1.88) and Labrador Retrievers (log OR = 1.09). In contrast, Dachshunds (log OR = −2.17), Jack Russell Terriers (log OR = −1.88), and Rhodesian Ridgebacks (log OR = −1.88) were rarely affected. This contrasted with the well-known predisposition of Dachshunds and Rhodesian Ridgebacks to oral and cutaneous melanocytic neoplasms. Further studies are needed to explain the underlying reasons for breed predisposition or “resistance” to the development of specific acral tumors and/or other sites. KW - canine KW - subungual KW - toe KW - tumor KW - inflammation KW - breed predisposition Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-242690 SN - 2306-7381 VL - 8 IS - 7 ER - TY - JOUR A1 - Lauruschkat, Chris D. A1 - Page, Lukas A1 - White, P. Lewis A1 - Etter, Sonja A1 - Davies, Helen E. A1 - Duckers, Jamie A1 - Ebel, Frank A1 - Schnack, Elisabeth A1 - Backx, Matthijs A1 - Dragan, Mariola A1 - Schlegel, Nicolas A1 - Kniemeyer, Olaf A1 - Brakhage, Axel A. A1 - Einsele, Hermann A1 - Loeffler, Juergen A1 - Wurster, Sebastian T1 - Development of a simple and robust whole blood assay with dual co-stimulation to quantify the release of T-cellular signature cytokines in response to Aspergillus fumigatus antigens JF - Journal of Fungi N2 - Deeper understanding of mold-induced cytokine signatures could promote advances in the diagnosis and treatment of invasive mycoses and mold-associated hypersensitivity syndromes. Currently, most T-cellular immunoassays in medical mycology require the isolation of mononuclear cells and have limited robustness and practicability, hampering their broader applicability in clinical practice. Therefore, we developed a simple, cost-efficient whole blood (WB) assay with dual α-CD28 and α-CD49d co-stimulation to quantify cytokine secretion in response to Aspergillus fumigatus antigens. Dual co-stimulation strongly enhanced A. fumigatus-induced release of T-cellular signature cytokines detectable by enzyme-linked immunosorbent assay (ELISA) or a multiplex cytokine assay. Furthermore, T-cell-dependent activation and cytokine response of innate immune cells was captured by the assay. The protocol consistently showed little technical variation and high robustness to pre-analytic delays of up to 8 h. Stimulation with an A. fumigatus lysate elicited at least 7-fold greater median concentrations of key T-helper cell signature cytokines, including IL-17 and the type 2 T-helper cell cytokines IL-4 and IL-5 in WB samples from patients with Aspergillus-associated lung pathologies versus patients with non-mold-related lung diseases, suggesting high discriminatory power of the assay. These results position WB-ELISA with dual co-stimulation as a simple, accurate, and robust immunoassay for translational applications, encouraging further evaluation as a platform to monitor host immunity to opportunistic pathogens. KW - immunoassay KW - biomarker KW - Aspergillus KW - cytokines KW - inflammation KW - adaptive immunity Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-241025 SN - 2309-608X VL - 7 IS - 6 ER - TY - JOUR A1 - Baum, Petra A1 - Toyka, Klaus V. A1 - Blüher, Matthias A1 - Kosacka, Joanna A1 - Nowicki, Marcin T1 - Inflammatory mechanisms in the pathophysiology of diabetic peripheral neuropathy (DN) — new aspects JF - International Journal of Molecular Sciences N2 - The pathogenesis of diabetic neuropathy is complex, and various pathogenic pathways have been proposed. A better understanding of the pathophysiology is warranted for developing novel therapeutic strategies. Here, we summarize recent evidence from experiments using animal models of type 1 and type 2 diabetes showing that low-grade intraneural inflammation is a facet of diabetic neuropathy. Our experimental data suggest that these mild inflammatory processes are a likely common terminal pathway in diabetic neuropathy associated with the degeneration of intraepidermal nerve fibers. In contrast to earlier reports claiming toxic effects of high-iron content, we found the opposite, i.e., nutritional iron deficiency caused low-grade inflammation and fiber degeneration while in normal or high non-heme iron nutrition no or only extremely mild inflammatory signs were identified in nerve tissue. Obesity and dyslipidemia also appear to trigger mild inflammation of peripheral nerves, associated with neuropathy even in the absence of overt diabetes mellitus. Our finding may be the experimental analog of recent observations identifying systemic proinflammatory activity in human sensorimotor diabetic neuropathy. In a rat model of type 1 diabetes, a mild neuropathy with inflammatory components could be induced by insulin treatment causing an abrupt reduction in HbA1c. This is in line with observations in patients with severe diabetes developing a small fiber neuropathy upon treatment-induced rapid HbA1c reduction. If the inflammatory pathogenesis could be further substantiated by data from human tissues and intervention studies, anti-inflammatory compounds with different modes of action may become candidates for the treatment or prevention of diabetic neuropathy. KW - diabetic neuropathy KW - pathogenesis KW - inflammation KW - iron KW - treatment-induced neuropathy in diabetes (TIND) Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-284556 SN - 1422-0067 VL - 22 IS - 19 ER - TY - JOUR A1 - Burkard, Natalie A1 - Meir, Michael A1 - Kannapin, Felix A1 - Otto, Christoph A1 - Petzke, Maximilian A1 - Germer, Christoph-Thomas A1 - Waschke, Jens A1 - Schlegel, Nicolas T1 - Desmoglein2 Regulates Claudin2 Expression by Sequestering PI-3-Kinase in Intestinal Epithelial Cells JF - Frontiers in Immunology N2 - Inflammation-induced reduction of intestinal desmosomal cadherin Desmoglein 2 (Dsg2) is linked to changes of tight junctions (TJ) leading to impaired intestinal epithelial barrier (IEB) function by undefined mechanisms. We characterized the interplay between loss of Dsg2 and upregulation of pore-forming TJ protein Claudin2. Intraperitoneal application of Dsg2-stablising Tandem peptide (TP) attenuated impaired IEB function, reduction of Dsg2 and increased Claudin2 in DSS-induced colitis in C57Bl/6 mice. TP blocked loss of Dsg2-mediated adhesion and upregulation of Claudin2 in Caco2 cells challenged with TNFα. In Dsg2-deficient Caco2 cells basal expression of Claudin2 was increased which was paralleled by reduced transepithelial electrical resistance and by augmented phosphorylation of AKT\(^{Ser473}\) under basal conditions. Inhibition of phosphoinositid-3-kinase proved that PI-3-kinase/AKT-signaling is critical to upregulate Claudin2. In immunostaining PI-3-kinase dissociated from Dsg2 under inflammatory conditions. Immunoprecipitations and proximity ligation assays confirmed a direct interaction of Dsg2 and PI-3-kinase which was abrogated following TNFα application. In summary, Dsg2 regulates Claudin2 expression by sequestering PI-3-kinase to the cell borders in intestinal epithelium. KW - Claudin2 KW - Dsg2 KW - inflammation KW - intestinal barrier KW - PI-3-kinase KW - inflammatory bowel disease KW - desmosome KW - tight junction Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-247059 SN - 1664-3224 VL - 12 ER - TY - JOUR A1 - Popp, Sandy A1 - Schmitt-Böhrer, Angelika A1 - Langer, Simon A1 - Hofmann, Ulrich A1 - Hommers, Leif A1 - Schuh, Kai A1 - Frantz, Stefan A1 - Lesch, Klaus-Peter A1 - Frey, Anna T1 - 5-HTT Deficiency in Male Mice Affects Healing and Behavior after Myocardial Infarction JF - Journal of Clinical Medicine N2 - Anxiety disorders and depression are common comorbidities in cardiac patients. Mice lacking the serotonin transporter (5-HTT) exhibit increased anxiety-like behavior. However, the role of 5-HTT deficiency on cardiac aging, and on healing and remodeling processes after myocardial infarction (MI), remains unclear. Cardiological evaluation of experimentally naïve male mice revealed a mild cardiac dysfunction in ≥4-month-old 5-HTT knockout (−/−) animals. Following induction of chronic cardiac dysfunction (CCD) by MI vs. sham operation 5-HTT−/− mice with infarct sizes >30% experienced 100% mortality, while 50% of 5-HTT+/− and 37% of 5-HTT+/+ animals with large MI survived the 8-week observation period. Surviving (sham and MI < 30%) 5-HTT−/− mutants displayed reduced exploratory activity and increased anxiety-like behavior in different approach-avoidance tasks. However, CCD failed to provoke a depressive-like behavioral response in either 5-Htt genotype. Mechanistic analyses were performed on mice 3 days post-MI. Electrocardiography, histology and FACS of inflammatory cells revealed no abnormalities. However, gene expression of inflammation-related cytokines (TGF-β, TNF-α, IL-6) and MMP-2, a protein involved in the breakdown of extracellular matrix, was significantly increased in 5-HTT−/− mice after MI. This study shows that 5-HTT deficiency leads to age-dependent cardiac dysfunction and disrupted early healing after MI probably due to alterations of inflammatory processes in mice. KW - chronic heart failure KW - myocardial infarction KW - serotonin transporter deficient mice KW - anxiety KW - depression KW - behavior KW - inflammation Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-242739 SN - 2077-0383 VL - 10 IS - 14 ER - TY - JOUR A1 - Heitmann, Johanna A1 - Frings, Verena G. A1 - Geier, Andreas A1 - Goebeler, Matthias A1 - Kerstan, Andreas T1 - Non-alcoholic fatty liver disease and psoriasis - is there a shared proinflammatory network? JF - Journal der Deutschen Dermatologischen Gesellschaft N2 - Psoriasis is an immune-mediated systemic inflammatory disease that is not limited to the skin but may be associated with arthritis, cardiovascular diseases, metabolic syndrome including diabetes and obesity and, as identified more recently, non-alcoholic fatty liver disease (NAFLD) that occurs in approximately 50 % of all patients with psoriasis. NAFLD is characterized by accumulation of fat in hepatocytes in the absence of excessive alcohol consumption. Over the last two decades, NAFLD has developed to the most common chronic liver disease with an estimated prevalence of 25 % in the Western population. NAFLD ranges from non-inflammatory or bland hepatic steatosis to inflammation of hepatic tissue (non-alcoholic steatohepatitis, NASH) and consecutive liver fibrosis. It is controversial whether the underlying systemic inflammation of psoriasis is contributing to development of NAFLD or if comorbid diseases such as obesity enhance NAFLD development. Recent findings indicate that cytokine-mediated inflammation through TNFα, interleukin (IL)-6 and IL-17 might be the common link between psoriasis and NAFLD. Considering the shared inflammatory pathways, IL-17 pharmacological blockade, which is already well-established for psoriasis, may be a promising strategy to treat both psoriasis and NAFLD. Therefore, early detection of NAFLD and a better understanding of its pathophysiology in the context of the systemic inflammation in psoriasis is important with regard to individualized treatment approaches. KW - psoriasis KW - fatty liver disease KW - inflammation Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-258424 VL - 19 IS - 4 ER - TY - JOUR A1 - Fröhlich, Matthias A1 - Serfling, Sebastian A1 - Higuchi, Takahiro A1 - Pomper, Martin G. A1 - Rowe, Steven P. A1 - Schmalzing, Marc A1 - Tony, Hans-Peter A1 - Gernert, Michael A1 - Strunz, Patrick-Pascal A1 - Portegys, Jan A1 - Schwaneck, Eva-Christina A1 - Gadeholt, Ottar A1 - Weich, Alexander A1 - Buck, Andreas K. A1 - Bley, Thorsten A. A1 - Guggenberger, Konstanze V. A1 - Werner, Rudolf A. T1 - Whole-Body [\(^{18}\)F]FDG PET/CT Can Alter Diagnosis in Patients with Suspected Rheumatic Disease JF - Diagnostics N2 - The 2-deoxy-d-[\(^{18}\)F]fluoro-D-glucose (FDG) positron emission tomography/computed tomography (PET/CT) is widely utilized to assess the vascular and articular inflammatory burden of patients with a suspected diagnosis of rheumatic disease. We aimed to elucidate the impact of [\(^{18}\)F]FDG PET/CT on change in initially suspected diagnosis in patients at the time of the scan. Thirty-four patients, who had undergone [\(^{18}\)F]FDG PET/CT, were enrolled and the initially suspected diagnosis prior to [18F]FDG PET/CT was compared to the final diagnosis. In addition, a semi-quantitative analysis including vessel wall-to-liver (VLR) and joint-to-liver (JLR) ratios was also conducted. Prior to [\(^{18}\)F]FDG PET/CT, 22/34 (64.7%) of patients did not have an established diagnosis, whereas in 7/34 (20.6%), polymyalgia rheumatica (PMR) was suspected, and in 5/34 (14.7%), giant cell arteritis (GCA) was suspected by the referring rheumatologists. After [\(^{18}\)F]FDG PET/CT, the diagnosis was GCA in 19/34 (55.9%), combined GCA and PMR (GCA + PMR) in 9/34 (26.5%) and PMR in the remaining 6/34 (17.6%). As such, [\(^{18}\)F]FDG PET/CT altered suspected diagnosis in 28/34 (82.4%), including in all unclear cases. VLR of patients whose final diagnosis was GCA tended to be significantly higher when compared to VLR in PMR (GCA, 1.01 ± 0.08 (95%CI, 0.95–1.1) vs. PMR, 0.92 ± 0.1 (95%CI, 0.85–0.99), p = 0.07), but not when compared to PMR + GCA (1.04 ± 0.14 (95%CI, 0.95–1.13), p = 1). JLR of individuals finally diagnosed with PMR (0.94 ± 0.16, (95%CI, 0.83–1.06)), however, was significantly increased relative to JLR in GCA (0.58 ± 0.04 (95%CI, 0.55–0.61)) and GCA + PMR (0.64 ± 0.09 (95%CI, 0.57–0.71); p < 0.0001, respectively). In individuals with a suspected diagnosis of rheumatic disease, an inflammatory-directed [\(^{18}\)F]FDG PET/CT can alter diagnosis in the majority of the cases, particularly in subjects who were referred because of diagnostic uncertainty. Semi-quantitative assessment may be helpful in establishing a final diagnosis of PMR, supporting the notion that a quantitative whole-body read-out may be useful in unclear cases. KW - giant cell arteritis KW - GCA KW - [18F]FDG PET/CT KW - vasculature KW - inflammation KW - polymyalgia rheumatica KW - PMR KW - vasculitis Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-250227 SN - 2075-4418 VL - 11 IS - 11 ER - TY - JOUR A1 - Morbach, Caroline A1 - Beyersdorf, Niklas A1 - Kerkau, Thomas A1 - Ramos, Gustavo A1 - Sahiti, Floran A1 - Albert, Judith A1 - Jahns, Roland A1 - Ertl, Georg A1 - Angermann, Christiane E. A1 - Frantz, Stefan A1 - Hofmann, Ulrich A1 - Störk, Stefan T1 - Adaptive anti-myocardial immune response following hospitalization for acute heart failure JF - ESC Heart Failure N2 - Aims It has been hypothesized that cardiac decompensation accompanying acute heart failure (AHF) episodes generates a pro-inflammatory environment boosting an adaptive immune response against myocardial antigens, thus contributing to progression of heart failure (HF) and poor prognosis. We assessed the prevalence of anti-myocardial autoantibodies (AMyA) as biomarkers reflecting adaptive immune responses in patients admitted to the hospital for AHF, followed the change in AMyA titres for 6 months after discharge, and evaluated their prognostic utility. Methods and results AMyA were determined in n = 47 patients, median age 71 (quartiles 60; 80) years, 23 (49%) female, and 24 (51%) with HF with preserved ejection fraction, from blood collected at baseline (time point of hospitalization) and at 6 month follow-up (visit F6). Patients were followed for 18 months (visit F18). The prevalence of AMyA increased from baseline (n = 21, 45%) to F6 (n = 36, 77%; P < 0.001). At F6, the prevalence of AMyA was higher in patients with HF with preserved ejection fraction (n = 21, 88%) compared with patients with reduced ejection fraction (n = 14, 61%; P = 0.036). During the subsequent 12 months after F6, that is up to F18, patients with newly developed AMyA at F6 had a higher risk for the combined endpoint of death or rehospitalization for HF (hazard ratio 4.79, 95% confidence interval 1.13–20.21; P = 0.033) compared with patients with persistent or without AMyA at F6. Conclusions Our results support the hypothesis that AHF may induce patterns of adaptive immune responses. More studies in larger populations and well-defined patient subgroups are needed to further clarify the role of the adaptive immune system in HF progression. KW - adaptive immune response KW - acute heart failure KW - anti-myocardial KW - autoantibody KW - inflammation Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-258907 VL - 8 IS - 4 ER - TY - JOUR A1 - Rajendran, Ranjithkumar A1 - Rajendran, Vinothkumar A1 - Giraldo-Velasquez, Mario A1 - Megalofonou, Fevronia-Foivi A1 - Gurski, Fynn A1 - Stadelmann, Christine A1 - Karnati, Srikanth A1 - Berghoff, Martin T1 - Oligodendrocyte-specific deletion of FGFR1 reduces cerebellar inflammation and neurodegeneration in MOG\(_{35-55}\)-induced EAE JF - International Journal of Molecular Sciences N2 - Multiple sclerosis (MS) is a chronic inflammatory and degenerative disease of the central nervous system (CNS). MS commonly affects the cerebellum causing acute and chronic symptoms. Cerebellar signs significantly contribute to clinical disability, and symptoms such as tremor, ataxia, and dysarthria are difficult to treat. Fibroblast growth factors (FGFs) and their receptors (FGFRs) are involved in demyelinating pathologies such as MS. In autopsy tissue from patients with MS, increased expression of FGF1, FGF2, FGF9, and FGFR1 was found in lesion areas. Recent research using mouse models has focused on regions such as the spinal cord, and data on the expression of FGF/FGFR in the cerebellum are not available. In recent EAE studies, we detected that oligodendrocyte-specific deletion of FGFRs results in a milder disease course, less cellular infiltrates, and reduced neurodegeneration in the spinal cord. The objective of this study was to characterize the role of FGFR1 in oligodendrocytes in the cerebellum. Conditional deletion of FGFR1 in oligodendrocytes (Fgfr1\(^{ind−/−}\) was achieved by tamoxifen application, EAE was induced using the MOG\(_{35-55}\) peptide. The cerebellum was analyzed by histology, immunohistochemistry, and western blot. At day 62 p.i., Fgfr1\(^{ind−/−}\) mice showed less myelin and axonal degeneration compared to FGFR1-competent mice. Infiltration of CD3(+) T cells, Mac3(+) cells, B220(+) B cells and IgG(+) plasma cells in cerebellar white matter lesions (WML) was less in Fgfr1\(^{ind−/−}\)mice. There were no effects on the number of OPC or mature oligodendrocytes in white matter lesion (WML). Expression of FGF2 and FGF9 associated with less myelin and axonal degeneration, and of the pro-inflammatory cytokines IL-1β, IL-6, and CD200 was downregulated in Fgfr1\(^{ind−/−}\) mice. The FGF/FGFR signaling protein pAkt, BDNF, and TrkB were increased in Fgfr1\(^{ind−/−}\) mice. These data suggest that cell-specific deletion of FGFR1 in oligodendrocytes has anti-inflammatory and neuroprotective effects in the cerebellum in the EAE disease model of MS. KW - FGFR1 KW - oligodendrocytes KW - demyelination KW - inflammation KW - cerebellum KW - EAE KW - MS Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-284296 SN - 1422-0067 VL - 22 IS - 17 ER - TY - JOUR A1 - Weiland, Judith A1 - Beez, Alexandra A1 - Westermaier, Thomas A1 - Kunze, Ekkehard A1 - Sirén, Anna-Leena A1 - Lilla, Nadine T1 - Neuroprotective strategies in aneurysmal subarachnoid hemorrhage (aSAH) JF - International Journal of Molecular Sciences N2 - Aneurysmal subarachnoid hemorrhage (aSAH) remains a disease with high mortality and morbidity. Since treating vasospasm has not inevitably led to an improvement in outcome, the actual emphasis is on finding neuroprotective therapies in the early phase following aSAH to prevent secondary brain injury in the later phase of disease. Within the early phase, neuroinflammation, thromboinflammation, disturbances in brain metabolism and early neuroprotective therapies directed against delayed cerebral ischemia (DCI) came into focus. Herein, the role of neuroinflammation, thromboinflammation and metabolism in aSAH is depicted. Potential neuroprotective strategies regarding neuroinflammation target microglia activation, metalloproteases, autophagy and the pathway via Toll-like receptor 4 (TLR4), high mobility group box 1 (HMGB1), NF-κB and finally the release of cytokines like TNFα or IL-1. Following the link to thromboinflammation, potential neuroprotective therapies try to target microthrombus formation, platelets and platelet receptors as well as clot clearance and immune cell infiltration. Potential neuroprotective strategies regarding metabolism try to re-balance the mismatch of energy need and supply following aSAH, for example, in restoring fuel to the TCA cycle or bypassing distinct energy pathways. Overall, this review addresses current neuroprotective strategies in aSAH, hopefully leading to future translational therapy options to prevent secondary brain injury. KW - subarachnoid hemorrhage (SAH) KW - inflammation KW - thromboinflammation KW - metabolism KW - neuroprotection KW - therapy Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-260755 SN - 1422-0067 VL - 22 IS - 11 ER - TY - JOUR A1 - Andelovic, Kristina A1 - Winter, Patrick A1 - Jakob, Peter Michael A1 - Bauer, Wolfgang Rudolf A1 - Herold, Volker A1 - Zernecke, Alma T1 - Evaluation of plaque characteristics and inflammation using magnetic resonance imaging JF - Biomedicines N2 - Atherosclerosis is an inflammatory disease of large and medium-sized arteries, characterized by the growth of atherosclerotic lesions (plaques). These plaques often develop at inner curvatures of arteries, branchpoints, and bifurcations, where the endothelial wall shear stress is low and oscillatory. In conjunction with other processes such as lipid deposition, biomechanical factors lead to local vascular inflammation and plaque growth. There is also evidence that low and oscillatory shear stress contribute to arterial remodeling, entailing a loss in arterial elasticity and, therefore, an increased pulse-wave velocity. Although altered shear stress profiles, elasticity and inflammation are closely intertwined and critical for plaque growth, preclinical and clinical investigations for atherosclerosis mostly focus on the investigation of one of these parameters only due to the experimental limitations. However, cardiovascular magnetic resonance imaging (MRI) has been demonstrated to be a potent tool which can be used to provide insights into a large range of biological parameters in one experimental session. It enables the evaluation of the dynamic process of atherosclerotic lesion formation without the need for harmful radiation. Flow-sensitive MRI provides the assessment of hemodynamic parameters such as wall shear stress and pulse wave velocity which may replace invasive and radiation-based techniques for imaging of the vascular function and the characterization of early plaque development. In combination with inflammation imaging, the analyses and correlations of these parameters could not only significantly advance basic preclinical investigations of atherosclerotic lesion formation and progression, but also the diagnostic clinical evaluation for early identification of high-risk plaques, which are prone to rupture. In this review, we summarize the key applications of magnetic resonance imaging for the evaluation of plaque characteristics through flow sensitive and morphological measurements. The simultaneous measurements of functional and structural parameters will further preclinical research on atherosclerosis and has the potential to fundamentally improve the detection of inflammation and vulnerable plaques in patients. KW - atherosclerosis KW - mouse models KW - wall shear stress KW - pulse wave velocity KW - arterial elasticity KW - inflammation KW - magnetic resonance imaging Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-228839 SN - 2227-9059 VL - 9 IS - 2 ER - TY - JOUR A1 - Ockermann, Philipp A1 - Headley, Laura A1 - Lizio, Rosario A1 - Hansmann, Jan T1 - A Review of the Properties of Anthocyanins and Their Influence on Factors Affecting Cardiometabolic and Cognitive Health JF - Nutrients N2 - The incidence of cardiovascular and metabolic diseases has increased over the last decades and is an important cause of death worldwide. An upcoming ingredient on the nutraceutical market are anthocyanins, a flavonoid subgroup, abundant mostly in berries and fruits. Epidemiological studies have suggested an association between anthocyanin intake and improved cardiovascular risk, type 2 diabetes and myocardial infarct. Clinical studies using anthocyanins have shown a significant decrease in inflammation markers and oxidative stress, a beneficial effect on vascular function and hyperlipidemia by decreasing low-density lipoprotein and increasing high-density lipoprotein. They have also shown a potential effect on glucose homeostasis and cognitive decline. This review summarizes the effects of anthocyanins in in-vitro, animal and human studies to give an overview of their application in medical prevention or as a dietary supplement. KW - anthocyanins KW - antioxidative KW - blood pressure KW - hyperlipidemia KW - diabetes KW - inflammation Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-245116 SN - 2072-6643 VL - 13 IS - 8 ER - TY - JOUR A1 - Graser, Stephanie A1 - Liedtke, Daniel A1 - Jakob, Franz T1 - TNAP as a new player in chronic inflammatory conditions and metabolism JF - International Journal of Molecular Sciences N2 - This review summarizes important information on the ectoenzyme tissue-nonspecific alkaline phosphatase (TNAP) and gives a brief insight into the symptoms, diagnostics, and treatment of the rare disease Hypophosphatasia (HPP), which is resulting from mutations in the TNAP encoding ALPL gene. We emphasize the role of TNAP beyond its well-known contribution to mineralization processes. Therefore, above all, the impact of the enzyme on central molecular processes in the nervous system and on inflammation is presented here. KW - TNAP KW - Hypophosphatasia KW - HPP KW - mineralization KW - nervous system KW - inflammation Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-258888 SN - 1422-0067 VL - 22 IS - 2 ER - TY - JOUR A1 - Budde, Heidi A1 - Hassoun, Roua A1 - Tangos, Melina A1 - Zhazykbayeva, Saltanat A1 - Herwig, Melissa A1 - Varatnitskaya, Marharyta A1 - Sieme, Marcel A1 - Delalat, Simin A1 - Sultana, Innas A1 - Kolijn, Detmar A1 - Gömöri, Kamilla A1 - Jarkas, Muhammad A1 - Lódi, Mária A1 - Jaquet, Kornelia A1 - Kovács, Árpád A1 - Mannherz, Hans Georg A1 - Sequeira, Vasco A1 - Mügge, Andreas A1 - Leichert, Lars I. A1 - Sossalla, Samuel A1 - Hamdani, Nazha T1 - The interplay between S-glutathionylation and phosphorylation of cardiac troponin I and myosin binding protein C in end-stage human failing hearts JF - Antioxidants N2 - Oxidative stress is defined as an imbalance between the antioxidant defense system and the production of reactive oxygen species (ROS). At low levels, ROS are involved in the regulation of redox signaling for cell protection. However, upon chronical increase in oxidative stress, cell damage occurs, due to protein, DNA and lipid oxidation. Here, we investigated the oxidative modifications of myofilament proteins, and their role in modulating cardiomyocyte function in end-stage human failing hearts. We found altered maximum Ca\(^{2+}\)-activated tension and Ca\(^{2+}\) sensitivity of force production of skinned single cardiomyocytes in end-stage human failing hearts compared to non-failing hearts, which was corrected upon treatment with reduced glutathione enzyme. This was accompanied by the increased oxidation of troponin I and myosin binding protein C, and decreased levels of protein kinases A (PKA)- and C (PKC)-mediated phosphorylation of both proteins. The Ca\(^{2+}\) sensitivity and maximal tension correlated strongly with the myofilament oxidation levels, hypo-phosphorylation, and oxidative stress parameters that were measured in all the samples. Furthermore, we detected elevated titin-based myocardial stiffness in HF myocytes, which was reversed by PKA and reduced glutathione enzyme treatment. Finally, many oxidative stress and inflammation parameters were significantly elevated in failing hearts compared to non-failing hearts, and corrected upon treatment with the anti-oxidant GSH enzyme. Here, we provide evidence that the altered mechanical properties of failing human cardiomyocytes are partially due to phosphorylation, S-glutathionylation, and the interplay between the two post-translational modifications, which contribute to the development of heart failure. KW - myofilament proteins KW - oxidative stress KW - inflammation KW - phosphorylation KW - S-glutathionylation Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-242701 SN - 2076-3921 VL - 10 IS - 7 ER - TY - THES A1 - Burow, Wera Tamara T1 - Die Rolle des CEACAM1-Moleküls bei der Entstehung von neurogener Entzündung in den Atemwegen T1 - The role of the CEACAM1 molecule in the development of neurogenic inflammation in the airways N2 - Neurogene Entzündung ist charakterisiert durch Vasodilatation, Plasmaextravasation und Leukozytenmigration. Im Zuge dieser Dissertationsarbeit konnte ein in vivo Versuchsmodell zur Quantifizierung neurogener Entzündungsreaktionen in den Atemwegen etabliert werden. Der bakterielle Bitterstoff Cycloheximid ist in der Lage, eine Erhöhung der Plasmaextravasation und Migration neutrophiler Granulozyten zu bewirken. Somit kann Cycloheximid nicht nur protektive Schutzreflexe auslösen, sondern führt auch lokal zu einer neurogenen Entzündungsreaktion. Das carcinoembryonic antigen-related cell adhesion molecule-1 (CEACAM1) ist an der Regulierung der endothelialen Barrierefunktion beteiligt. Die Versuche zeigen bei CC1-/--Mäusen eine Verminderung der basalen Permeabilität in trachealen postkapillären Venolen. Nach Stimulation mit Cycloheximid zeigen CC1-/--Mäuse im Vergleich mit WT-Mäusen eine verminderte Plasmaextravasation in bronchialen postkapillären Venolen. Auch die Permeabilität des Endothels für neutrophile Granulozyten scheint durch CEACAM1-Defizienz in trachealen und bronchialen Venolen herabgesetzt zu werden. Die Anwesenheit des CEACAM1-Moleküls verursacht offenbar eine verminderte Stabilität der endothelialen Barriere in postkapillären Venolen der Atemwege. Diese Ergebnisse zeigen eine gegenteilige Funktion von CEACAM1 in postkapillären Venolen der Atemwege im Vergleich mit großen, herznahen Blutgefäßen. Des Weiteren scheint sich die Rolle von CEACAM1 in der Entstehung von akuten und chronischen Entzündungsreaktionen zu unterscheiden. Das in dieser Arbeit etablierte Versuchsmodell stellt eine Möglichkeit dar, neurogene Entzündungsreaktionen als Reaktion auf verschiedene gustatorische Stimulanzien zu testen und zu quantifizieren. N2 - Neurogenic inflammation is characterized by vasodilatation, plasma extravasation and leukocyte recruitment. This thesis presents an in vivo model for quantification of neurogenic inflammatory reactions in the airways. Application of the bacterial bitter substance Cycloheximide results in an increased plasma extravasation and migration of neutrophil granulocytes. Cycloheximide not only triggers protective reflexes as it has been previously shown but it also causes local neurogenic inflammatory responses. The carcinoembryonic antigen-related cell adhesion molecule-1 (CEACAM1) is known to regulate endothelial barrier function. Experiments show a decrease in basal vascular permeability in CEACAM1 knock-out mice (CC1-/-) in tracheal postcapillary venules. After stimulation with Cycloheximide CC1-/--mice exhibit a decreased plasma extravasation in bronchial postcapillary venules compared to wild-type mice. The permeability of the endothelium to neutrophil granulocytes is also decreased in tracheal and bronchial postcapillary venules of CC1-/--mice. Expression of the CEACAM1 molecule leads to a reduced stability of the endothelial barrier in postcapillary venules of the respiratory tract. These results show an opposite function of CEACAM1 in postcapillary airway venules compared to larger vessels. Furthermore, the role of CEACAM1 appears to be different in the development of acute and chronic inflammatory reactions. The established in vivo model offers a possibility to quantify neurogenic inflammation in response to various gustatory stimuli. KW - Entzündung KW - Atemwege KW - neurogenic KW - inflammation KW - CEACAM1 KW - Cycloheximide KW - mouse model Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-209331 ER - TY - JOUR A1 - Schäfer, Sarah A1 - Zernecke, Alma T1 - CD8\(^+\) T cells in atherosclerosis JF - Cells N2 - Atherosclerotic lesions are populated by cells of the innate and adaptive immune system, including CD8\(^+\) T cells. The CD8\(^+\) T cell infiltrate has recently been characterized in mouse and human atherosclerosis and revealed activated, cytotoxic, and possibly dysfunctional and exhausted cell phenotypes. In mouse models of atherosclerosis, antibody-mediated depletion of CD8\(^+\) T cells ameliorates atherosclerosis. CD8\(^+\) T cells control monopoiesis and macrophage accumulation in early atherosclerosis. In addition, CD8\(^+\) T cells exert cytotoxic functions in atherosclerotic plaques and contribute to macrophage cell death and necrotic core formation. CD8\(^+\) T cell activation may be antigen-specific, and epitopes of atherosclerosis-relevant antigens may be targets of CD8\(^+\) T cells and their cytotoxic activity. CD8\(^+\) T cell functions are tightly controlled by costimulatory and coinhibitory immune checkpoints. Subsets of regulatory CD25\(^+\)CD8\(^+\) T cells with immunosuppressive functions can inhibit atherosclerosis. Importantly, local cytotoxic CD8\(^+\) T cell responses may trigger endothelial damage and plaque erosion in acute coronary syndromes. Understanding the complex role of CD8\(^+\) T cells in atherosclerosis may pave the way for defining novel treatment approaches in atherosclerosis. In this review article, we discuss these aspects, highlighting the emerging and critical role of CD8\(^+\) T cells in atherosclerosis. KW - atherosclerosis KW - CD8\(^+\) T cells KW - inflammation KW - cytotoxic T cells KW - single cell RNA sequencing KW - checkpoint inhibitors KW - immunotherapy Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-220170 SN - 2073-4409 VL - 10 IS - 1 ER - TY - JOUR A1 - Frischholz, Sebastian A1 - Berberich, Oliver A1 - Böck, Thomas A1 - Meffert, Rainer H. A1 - Blunk, Torsten T1 - Resveratrol counteracts IL‐1β‐mediated impairment of extracellular matrix deposition in 3D articular chondrocyte constructs JF - Journal of Tissue Engineering and Regenerative Medicine N2 - When aiming at cell‐based therapies in osteoarthritis (OA), proinflammatory conditions mediated by cytokines such as IL‐1β need to be considered. In recent studies, the phytoalexin resveratrol (RSV) has exhibited potent anti‐inflammatory properties. However, long‐term effects on 3D cartilaginous constructs under inflammatory conditions with regard to tissue quality, especially extracellular matrix (ECM) composition, have remained unexplored. Therefore, we employed long‐term model cultures for cell‐based therapies in an in vitro OA environment and evaluated effects of RSV. Pellet constructs made from expanded porcine articular chondrocytes were cultured with either IL‐1β (1–10 ng/ml) or RSV (50 μM) alone, or a cotreatment with both agents. Treatments were applied for 14 days, either directly after pellet formation or after a preculture period of 7 days. Culture with IL‐1β (10 ng/ml) decreased pellet size and DNA amount and severely compromised glycosaminoglycan (GAG) and collagen content. Cotreatment with RSV distinctly counteracted the proinflammatory catabolism and led to partial rescue of the ECM composition in both culture systems, with especially strong effects on GAG. Marked MMP13 expression was detected in IL‐1β‐treated pellets, but none upon RSV cotreatment. Expression of collagen type I was increased upon IL‐1β treatment and still observed when adding RSV, whereas collagen type X, indicating hypertrophy, was detected exclusively in pellets treated with RSV alone. In conclusion, RSV can counteract IL‐1β‐mediated degradation and distinctly improve cartilaginous ECM deposition in 3D long‐term inflammatory cultures. Nevertheless, potential hypertrophic effects should be taken into account when considering RSV as cotreatment for articular cartilage repair techniques. KW - articular chondrocytes KW - cartilage KW - cell‐based therapy KW - extracellular matrix KW - IL‐1β KW - inflammation KW - osteoarthritis KW - resveratrol Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-215471 VL - 14 IS - 7 SP - 897 EP - 908 ER - TY - JOUR A1 - Oelschlaegel, Diana A1 - Weiss Sadan, Tommy A1 - Salpeter, Seth A1 - Krug, Sebastian A1 - Blum, Galia A1 - Schmitz, Werner A1 - Schulze, Almut A1 - Michl, Patrick T1 - Cathepsin inhibition modulates metabolism and polarization of tumor-associated macrophages JF - Cancers N2 - Stroma-infiltrating immune cells, such as tumor-associated macrophages (TAM), play an important role in regulating tumor progression and chemoresistance. These effects are mostly conveyed by secreted mediators, among them several cathepsin proteases. In addition, increasing evidence suggests that stroma-infiltrating immune cells are able to induce profound metabolic changes within the tumor microenvironment. In this study, we aimed to characterize the impact of cathepsins in maintaining the TAM phenotype in more detail. For this purpose, we investigated the molecular effects of pharmacological cathepsin inhibition on the viability and polarization of human primary macrophages as well as its metabolic consequences. Pharmacological inhibition of cathepsins B, L, and S using a novel inhibitor, GB111-NH\(_2\), led to changes in cellular recycling processes characterized by an increased expression of autophagy- and lysosome-associated marker genes and reduced adenosine triphosphate (ATP) content. Decreased cathepsin activity in primary macrophages further led to distinct changes in fatty acid metabolites associated with increased expression of key modulators of fatty acid metabolism, such as fatty acid synthase (FASN) and acid ceramidase (ASAH1). The altered fatty acid profile was associated with an increased synthesis of the pro-inflammatory prostaglandin PGE\(_2\), which correlated with the upregulation of numerous NF\(_k\)B-dependent pro-inflammatory mediators, including interleukin-1 (IL-1), interleukin-6 (IL-6), C-C motif chemokine ligand 2 (CCL2), and tumor necrosis factor-alpha (TNFα). Our data indicate a novel link between cathepsin activity and metabolic reprogramming in macrophages, demonstrated by a profound impact on autophagy and fatty acid metabolism, which facilitates a pro-inflammatory micromilieu generally associated with enhanced tumor elimination. These results provide a strong rationale for therapeutic cathepsin inhibition to overcome the tumor-promoting effects of the immune-evasive tumor micromilieu. KW - cathepsin KW - activity-based probes KW - tumor-associated macrophage KW - autophagy KW - lysosome KW - lipid metabolism KW - inflammation Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-213040 SN - 2072-6694 VL - 12 IS - 9 ER - TY - JOUR A1 - Mahmood, Zafar A1 - Schmalzing, Marc A1 - Dörner, Thomas A1 - Tony, Hans-Peter A1 - Muhammad, Khalid T1 - Therapeutic Cytokine Inhibition Modulates Activation and Homing Receptors of Peripheral Memory B Cell Subsets in Rheumatoid Arthritis Patients JF - Frontiers in Immunology N2 - Memory B cells have known to play an important role in the pathogenesis of rheumatoid arthritis (RA). With the emergence of B cell-targeted therapies, the modulation of memory B cells appears to be a key therapeutic target. Human peripheral memory B cells can be distinguished based on the phenotypic expression of CD27 and IgD, characterizing the three major B cell subpopulations: CD27+IgD+ pre-switch, CD27+IgD- post-switch, and CD27-IgD- double-negative memory B cells. We evaluated different memory cell populations for activation markers (CD95 and Ki-67) and chemokine receptors (CXCR3 and 4) expressing B cells in active RA, as well as under IL6-R blockade by tocilizumab (TCZ) and TNF-α blockade by adalimumab (ADA). Memory B cells were phenotypically analyzed from RA patients at baseline, week 12, and week 24 under TCZ or ADA treatment, respectively. Using flow cytometry, surface expression of CD95, intracellular Ki-67, and surface expressions of CXCR3 and CXCR4 were determined. Compared with healthy donors (n = 40), the phenotypic analysis of RA patients (n = 80) demonstrated that all three types of memory B cells were activated in RA patients. Surface and intracellular staining of B cells showed a significantly higher percentage of CD95+ (p < 0.0001) and Ki-67+ (p < 0.0001) cells, with numerically altered CXCR3+ and CXCR4+ cells in RA. CD95 and Ki-67 expressions were highest in post-switch memory B cells, whereas CD19+CXCR3+ and CD19+CXCR4+ expressing cells were substantially higher in the pre-switch compartment. In all subsets of the memory B cells, in vivo IL-6R, and TNF-α blockade significantly reduced the enhanced expressions of CD95 and Ki-67. Based on our findings, we conclude that the three major peripheral memory B cell populations, pre-, post-switch, and double-negative B cells, are activated in RA, demonstrating enhanced CD95 and Ki-67 expressions, and varied expression of CXCR3 and CXCR4 chemokine receptors when compared with healthy individuals. This activation can be efficaciously modulated under cytokine inhibition in vivo. KW - B cells KW - inflammation KW - adalimumab KW - tocilizumab (IL-6 inhibitor) KW - memory B cells KW - rheumatoid arhritis Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-212380 SN - 1664-3224 VL - 11 ER - TY - JOUR A1 - Gabbert, Lydia A1 - Dilling, Christina A1 - Meybohm, Patrick A1 - Burek, Malgorzata T1 - Deletion of Protocadherin Gamma C3 Induces Phenotypic and Functional Changes in Brain Microvascular Endothelial Cells In Vitro JF - Frontiers in Pharmacology N2 - Inflammation of the central nervous system (CNS) is associated with diseases such as multiple sclerosis, stroke and neurodegenerative diseases. Compromised integrity of the blood-brain barrier (BBB) and increased migration of immune cells into the CNS are the main characteristics of brain inflammation. Clustered protocadherins (Pcdhs) belong to a large family of cadherin-related molecules. Pcdhs are highly expressed in the CNS in neurons, astrocytes, pericytes and epithelial cells of the choroid plexus and, as we have recently demonstrated, in brain microvascular endothelial cells (BMECs). Knockout of a member of the Pcdh subfamily, PcdhgC3, resulted in significant changes in the barrier integrity of BMECs. Here we characterized the endothelial PcdhgC3 knockout (KO) cells using paracellular permeability measurements, proliferation assay, wound healing assay, inhibition of signaling pathways, oxygen/glucose deprivation (OGD) and a pro-inflammatory cytokine tumor necrosis factor alpha (TNFα) treatment. PcdhgC3 KO showed an increased paracellular permeability, a faster proliferation rate, an altered expression of efflux pumps, transporters, cellular receptors, signaling and inflammatory molecules. Serum starvation led to significantly higher phosphorylation of extracellular signal-regulated kinases (Erk) in KO cells, while no changes in phosphorylated Akt kinase levels were found. PcdhgC3 KO cells migrated faster in the wound healing assay and this migration was significantly inhibited by respective inhibitors of the MAPK-, β-catenin/Wnt-, mTOR- signaling pathways (SL327, XAV939, or Torin 2). PcdhgC3 KO cells responded stronger to OGD and TNFα by significantly higher induction of interleukin 6 mRNA than wild type cells. These results suggest that PcdhgC3 is involved in the regulation of major signaling pathways and the inflammatory response of BMECs. KW - blood-brain barrier KW - protocadherin gamma C3 KW - inflammation KW - oxygen/glucose deprivation KW - stroke KW - tumor necrosis factor-α KW - proliferation Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-219828 SN - 1663-9812 VL - 11 ER - TY - JOUR A1 - Notz, Quirin A1 - Schmalzing, Marc A1 - Wedekink, Florian A1 - Schlesinger, Tobias A1 - Gernert, Michael A1 - Herrmann, Johannes A1 - Sorger, Lena A1 - Weismann, Dirk A1 - Schmid, Benedikt A1 - Sitter, Magdalena A1 - Schlegel, Nicolas A1 - Kranke, Peter A1 - Wischhusen, Jörg A1 - Meybohm, Patrick A1 - Lotz, Christopher T1 - Pro- and Anti-Inflammatory Responses in Severe COVID-19-Induced Acute Respiratory Distress Syndrome—An Observational Pilot Study JF - Frontiers in Immunology N2 - Objectives The severity of Coronavirus Disease 2019 (COVID-19) is largely determined by the immune response. First studies indicate altered lymphocyte counts and function. However, interactions of pro- and anti-inflammatory mechanisms remain elusive. In the current study we characterized the immune responses in patients suffering from severe COVID-19-induced acute respiratory distress syndrome (ARDS). Methods This was a single-center retrospective study in patients admitted to the intensive care unit (ICU) with confirmed COVID-19 between March 14th and May 28th 2020 (n = 39). Longitudinal data were collected within routine clinical care, including flow-cytometry of lymphocyte subsets, cytokine analysis and growth differentiation factor 15 (GDF-15). Antibody responses against the receptor binding domain (RBD) of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Spike protein were analyzed. Results All patients suffered from severe ARDS, 30.8% died. Interleukin (IL)-6 was massively elevated at every time-point. The anti-inflammatory cytokine IL-10 was concomitantly upregulated with IL-6. The cellular response was characterized by lymphocytopenia with low counts of CD8+ T cells, natural killer (NK) and naïve T helper cells. CD8+ T and NK cells recovered after 8 to 14 days. The B cell system was largely unimpeded. This coincided with a slight increase in anti-SARS-CoV-2-Spike-RBD immunoglobulin (Ig) G and a decrease in anti-SARS-CoV-2-Spike-RBD IgM. GDF-15 levels were elevated throughout ICU treatment. Conclusions Massively elevated levels of IL-6 and a delayed cytotoxic immune defense characterized severe COVID-19-induced ARDS. The B cell response and antibody production were largely unimpeded. No obvious imbalance of pro- and anti-inflammatory mechanisms was observed, with elevated GDF-15 levels suggesting increased tissue resilience. KW - Coronavirus Disease 2019 KW - acute respiratory distress syndrome KW - Severe Acute Respiratory Syndrome Coronavirus 2 KW - cytokines KW - inflammation KW - growth differentiation factor 15 KW - immune response Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-212815 SN - 1664-3224 VL - 11 ER - TY - JOUR A1 - Kleefeldt, Florian A1 - Bömmel, Heike A1 - Broede, Britta A1 - Thomsen, Michael A1 - Pfeiffer, Verena A1 - Wörsdörfer, Philipp A1 - Karnati, Srikanth A1 - Wagner, Nicole A1 - Rueckschloss, Uwe A1 - Ergün, Süleyman T1 - Aging‐related carcinoembryonic antigen‐related cell adhesion molecule 1 signaling promotes vascular dysfunction JF - Aging Cell N2 - Aging is an independent risk factor for cardiovascular diseases and therefore of particular interest for the prevention of cardiovascular events. However, the mechanisms underlying vascular aging are not well understood. Since carcinoembryonic antigen‐related cell adhesion molecule 1 (CEACAM1) is crucially involved in vascular homeostasis, we sought to identify the role of CEACAM1 in vascular aging. Using human internal thoracic artery and murine aorta, we show that CEACAM1 is upregulated in the course of vascular aging. Further analyses demonstrated that TNF‐α is CEACAM1‐dependently upregulated in the aging vasculature. Vice versa, TNF‐α induces CEACAM1 expression. This results in a feed‐forward loop in the aging vasculature that maintains a chronic pro‐inflammatory milieu. Furthermore, we demonstrate that age‐associated vascular alterations, that is, increased oxidative stress and vascular fibrosis, due to increased medial collagen deposition crucially depend on the presence of CEACAM1. Additionally, age‐dependent upregulation of vascular CEACAM1 expression contributes to endothelial barrier impairment, putatively via increased VEGF/VEGFR‐2 signaling. Consequently, aging‐related upregulation of vascular CEACAM1 expression results in endothelial dysfunction that may promote atherosclerotic plaque formation in the presence of additional risk factors. Our data suggest that CEACAM1 might represent an attractive target in order to delay physiological aging and therefore the transition to vascular disorders such as atherosclerosis. KW - aging KW - anti‐aging KW - cytokines KW - inflammation KW - mouse KW - reactive oxygen species Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-201231 VL - 2019 IS - 18 ER - TY - JOUR A1 - Glaser, Kirsten A1 - Gradzka-Luczewska, Anna A1 - Szymankiewicz-Breborowicz, Marta A1 - Kawczynska-Leda, Natalia A1 - Henrich, Birgit A1 - Waaga-Gasser, Ana Maria A1 - Speer, Christian P. T1 - Perinatal ureaplasma exposure is associated with increased risk of late onset sepsis and imbalanced inflammation in preterm infants and may add to lung injury JF - Frontiers in Cellular and Infection Microbiology N2 - Background: Controversy remains concerning the impact of Ureaplasma on preterm neonatal morbidity. Methods: Prospective single-center study in very low birth weight infants <30 weeks' gestation. Cord blood and initial nasopharyngeal swabs were screened for Ureaplasma parvum and U. urealyticum using culture technique and polymerase chain reaction. Neonatal outcomes were followed until death or discharge. Multi-analyte immunoassay provided cord blood levels of inflammatory markers. Using multivariate regression analyses, perinatal Ureaplasma exposure was evaluated as risk factor for the development of bronchopulmonary dysplasia (BPD), other neonatal morbidities until discharge and systemic inflammation at admission. Results: 40/103 (39%) infants were positive for Ureaplasma in one or both specimens, with U. parvum being the predominant species. While exposure to Ureaplasma alone was not associated with BPD, we found an increased risk of BPD in Ureaplasma-positive infants ventilated ≥5 days (OR 1.64; 95% CI 0.12–22.98; p = 0.009). Presence of Ureaplasma was associated with a 7-fold risk of late onset sepsis (LOS) (95% CI 1.80–27.39; p = 0.014). Moreover, Ureaplasma-positive infants had higher I/T ratios (b 0.39; 95% CI 0.08–0.71; p = 0.014), increased levels of interleukin (IL)-17 (b 0.16; 95% CI 0.02–0.30; p = 0.025) and matrix metalloproteinase 8 (b 0.77; 95% CI 0.10–1.44; p = 0.020), decreased levels of IL-10 (b −0.77; 95% CI −1.58 to −0.01; p = 0.043) and increased ratios of Tumor necrosis factor-α, IL-8, and IL-17 to anti-inflammatory IL-10 (p = 0.003, p = 0.012, p < 0.001). Conclusions: Positive Ureaplasma screening was not associated with BPD. However, exposure contributed to BPD in infants ventilated ≥5 days and conferred an increased risk of LOS and imbalanced inflammatory cytokine responses. KW - Ureaplasma parvum KW - Ureaplasma urealyticum KW - preterm infants KW - VLBW KW - bronchopulmonary dysplasia KW - late onset sepsis KW - neonatal outcome KW - inflammation Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-201270 VL - 9 IS - 68 ER - TY - JOUR A1 - Muenstermann, Marcel A1 - Strobel, Lea A1 - Klos, Andreas A1 - Wetsel, Rick A. A1 - Woodruff, Trent M. A1 - Köhl, Jörg A1 - Johswich, Kay O. T1 - Distinct roles of the anaphylatoxin receptors C3aR, C5aR1 and C5aR2 in experimental meningococcal infections JF - Virulence N2 - The complement system is pivotal in the defense against invasive disease caused by Neisseria meningitidis (Nme, meningococcus), particularly via the membrane attack complex. Complement activation liberates the anaphylatoxins C3a and C5a, which activate three distinct G-protein coupled receptors, C3aR, C5aR1 and C5aR2 (anaphylatoxin receptors, ATRs). We recently discovered that C5aR1 exacerbates the course of the disease, revealing a downside of complement in Nme sepsis. Here, we compared the roles of all three ATRs during mouse nasal colonization, intraperitoneal infection and human whole blood infection with Nme. Deficiency of complement or ATRs did not alter nasal colonization, but significantly affected invasive disease: Compared to WT mice, the disease was aggravated in C3ar\(^{-/-}\) mice, whereas C5ar1\(^{-/-}\) and C5ar2\(^{-/-}\) mice showed increased resistance to meningococcal sepsis. Surprisingly, deletion of either of the ATRs resulted in lower cytokine/chemokine responses, irrespective of the different susceptibilities of the mice. This was similar in ex vivo human whole blood infection using ATR inhibitors. Neutrophil responses to Nme were reduced in C5ar1\(^{-/-}\) mouse blood. Upon stimulation with C5a plus Nme, mouse macrophages displayed reduced phosphorylation of ERK1/2, when C5aR1 or C5aR2 were ablated or inhibited, suggesting that both C5a-receptors prime an initial macrophage response to Nme. Finally, in vivo blockade of C5aR1 alone (PMX205) or along with C5aR2 (A8\(^{Δ71−73}\)) resulted in ameliorated disease, whereas neither antagonizing C3aR (SB290157) nor its activation with a “super-agonist” peptide (WWGKKYRASKLGLAR) demonstrated a benefit. Thus, C5aR1 and C5aR2 augment disease pathology and are interesting targets for treatment, whereas C3aR is protective in experimental meningococcal sepsis. KW - inflammation KW - C3a KW - C5a KW - C3aR KW - C5aR1 KW - C5aR2 KW - meningococcal disease KW - sepsis Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-200496 VL - 10 IS - 1 ER - TY - JOUR A1 - Rapa, Shara Francesca A1 - Di Iorio, Biagio Raffaele A1 - Campiglia, Pietro A1 - Heidland, August A1 - Marzocco, Stefania T1 - Inflammation and oxidative stress in chronic kidney disease — Potential therapeutic role of minerals, vitamins and plant-derived metabolites JF - International Journal of Molecular Sciences N2 - Chronic kidney disease (CKD) is a debilitating pathology with various causal factors, culminating in end stage renal disease (ESRD) requiring dialysis or kidney transplantation. The progression of CKD is closely associated with systemic inflammation and oxidative stress, which are responsible for the manifestation of numerous complications such as malnutrition, atherosclerosis, coronary artery calcification, heart failure, anemia and mineral and bone disorders, as well as enhanced cardiovascular mortality. In addition to conventional therapy with anti-inflammatory and antioxidative agents, growing evidence has indicated that certain minerals, vitamins and plant-derived metabolites exhibit beneficial effects in these disturbances. In the current work, we review the anti-inflammatory and antioxidant properties of various agents which could be of potential benefit in CKD/ESRD. However, the related studies were limited due to small sample sizes and short-term follow-up in many trials. Therefore, studies of several anti-inflammatory and antioxidant agents with long-term follow-ups are necessary. KW - chronic kidney disease (CKD) KW - inflammation KW - oxidative stress KW - uremic toxins KW - minerals KW - vitamins KW - plant-derived metabolites Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-284998 SN - 1422-0067 VL - 21 IS - 1 ER - TY - JOUR A1 - Scheurer, Mario Joachim Johannes A1 - Brands, Roman Camillus A1 - El-Mesery, Mohamed A1 - Hartmann, Stefan A1 - Müller-Richter, Urs Dietmar Achim A1 - Kübler, Alexander Christian A1 - Seher, Axel T1 - The selection of NFκB inhibitors to block inflammation and induce sensitisation to FasL-induced apoptosis in HNSCC cell lines is critical for their use as a prospective cancer therapy JF - International Journal of Molecular Science N2 - Inflammation is a central aspect of tumour biology and can contribute significantly to both the origination and progression of tumours. The NFκB pathway is one of the most important signal transduction pathways in inflammation and is, therefore, an excellent target for cancer therapy. In this work, we examined the influence of four NFκB inhibitors — Cortisol, MLN4924, QNZ and TPCA1 — on proliferation, inflammation and sensitisation to apoptosis mediated by the death ligand FasL in the HNSCC cell lines PCI1, PCI9, PCI13, PCI52 and SCC25 and in the human dermal keratinocyte cell line HaCaT. We found that the selection of the inhibitor is critical to ensure that cells do not respond by inducing counteracting activities in the context of cancer therapy, e.g., the extreme IL-8 induction mediated by MLN4924 or FasL resistance mediated by Cortisol. However, TPCA1 was qualified by this in vitro study as an excellent therapeutic mediator in HNSCC by four positive qualities: (1) proliferation was inhibited at low μM-range concentrations; (2) TNFα-induced IL-8 secretion was blocked; (3) HNSCC cells were sensitized to TNFα-induced cell death; and (4) FasL-mediated apoptosis was not disrupted. KW - HNSCC KW - NFκB KW - inhibitor KW - TPCA1 KW - apoptosis KW - inflammation KW - TNFα KW - FasL Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-201524 SN - 1422-0067 VL - 20 IS - 6 ER - TY - JOUR A1 - Jahn, Daniel A1 - Dorbath, Donata A1 - Kircher, Stefan A1 - Nier, Anika A1 - Bergheim, Ina A1 - Lenaerts, Kaatje A1 - Hermanns, Heike M. A1 - Geier, Andreas T1 - Beneficial effects of vitamin D treatment in an obese mouse model of non-alcoholic steatohepatitis JF - Nutrients N2 - Serum vitamin D levels negatively correlate with obesity and associated disorders such as non-alcoholic steatohepatitis (NASH). However, the mechanisms linking low vitamin D (VD) status to disease progression are not completely understood. In this study, we analyzed the effect of VD treatment on NASH in mice. C57BL6/J mice were fed a high-fat/high-sugar diet (HFSD) containing low amounts of VD for 16 weeks to induce obesity, NASH and liver fibrosis. The effects of preventive and interventional VD treatment were studied on the level of liver histology and hepatic/intestinal gene expression. Interestingly, preventive and to a lesser extent also interventional VD treatment resulted in improvements of liver histology. This included a significant decrease of steatosis, a trend towards lower non-alcoholic fatty liver disease (NAFLD) activity score and a slight non-significant decrease of fibrosis in the preventive treatment group. In line with these changes, preventive VD treatment reduced the hepatic expression of lipogenic, inflammatory and pro-fibrotic genes. Notably, these beneficial effects occurred in conjunction with a reduction of intestinal inflammation. Together, our observations suggest that timely initiation of VD supplementation (preventive vs. interventional) is a critical determinant of treatment outcome in NASH. In the applied animal model, the improvements of liver histology occurred in conjunction with reduced inflammation in the gut, suggesting a potential relevance of vitamin D as a therapeutic agent acting on the gut–liver axis. KW - vitamin D KW - obesity KW - NAFLD KW - NASH KW - inflammation KW - intestine KW - gut–liver axis Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-177222 VL - 11 IS - 1 ER - TY - JOUR A1 - Werner, Rudolf A1 - Wakabayashi, Hiroshi A1 - Bauer, Jochen A1 - Schütz, Claudia A1 - Zechmeister, Christina A1 - Hayakawa, Nobuyuki A1 - Javadi, Mehrbod S. A1 - Lapa, Constantin A1 - Jahns, Roland A1 - Ergün, Süleyman A1 - Jahns, Valerie A1 - Higuchi, Takahiro T1 - Longitudinal \(^{18}\)F-FDG PET imaging in a Rat Model of Autoimmune Myocarditis JF - European Heart Journal Cardiovascular Imaging N2 - Aims: Although mortality rate is very high, diagnosis of acute myocarditis remains challenging with conventional tests. We aimed to elucidate the potential role of longitudinal 2-Deoxy-2-\(^{18}\)F-fluoro-D-glucose (\(^{18}\)F-FDG) positron emission tomography (PET) inflammation monitoring in a rat model of experimental autoimmune myocarditis. Methods and results: Autoimmune myocarditis was induced in Lewis rats by immunizing with porcine cardiac myosin emulsified in complete Freund’s adjuvant. Time course of disease was assessed by longitudinal \(^{18}\)F-FDG PET imaging. A correlative analysis between in- and ex vivo \(^{18}\)F-FDG signalling and macrophage infiltration using CD68 staining was conducted. Finally, immunohistochemistry analysis of the cell-adhesion markers CD34 and CD44 was performed at different disease stages determined by longitudinal \(^{18}\)F-FDG PET imaging. After immunization, myocarditis rats revealed a temporal increase in 18F-FDG uptake (peaked at week 3), which was followed by a rapid decline thereafter. Localization of CD68 positive cells was well correlated with in vivo \(^{18}\)F-FDG PET signalling (R\(^2\) = 0.92) as well as with ex vivo 18F-FDG autoradiography (R\(^2\) = 0.9, P < 0.001, respectively). CD44 positivity was primarily observed at tissue samples obtained at acute phase (i.e. at peak 18F-FDG uptake), while CD34-positive staining areas were predominantly identified in samples harvested at both sub-acute and chronic phases (i.e. at \(^{18}\)F-FDG decrease). Conclusion: \(^{18}\)F-FDG PET imaging can provide non-invasive serial monitoring of cardiac inflammation in a rat model of acute myocarditis. KW - positron emission tomography KW - Myokarditis KW - myocarditis KW - inflammation KW - 18F-FDG KW - PET KW - personalized treatment Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-165601 SN - 2047-2404 ER - TY - JOUR A1 - Saint Fleur-Lominy, Shella A1 - Maus, Mate A1 - Vaeth, Martin A1 - Lange, Ingo A1 - Zee, Isabelle A1 - Suh, David A1 - Liu, Cynthia A1 - Wu, Xiaojun A1 - Tikhonova, Anastasia A1 - Aifantis, Iannis A1 - Feske, Stefan T1 - STIM1 and STIM2 Mediate Cancer-Induced Inflammation in T Cell Acute Lymphoblastic Leukemia JF - Cell Reports N2 - T cell acute lymphoblastic leukemia (T-ALL) is commonly associated with activating mutations in the NOTCH1 pathway. Recent reports have shown a link between NOTCH1 signaling and intracellular Ca2+ homeostasis in T-ALL. Here, we investigate the role of store-operated Ca2+ entry (SOCE) mediated by the Ca2+ channel ORAI1 and its activators STIM1 and STIM2 in T-ALL. Deletion of STIM1 and STIM2 in leukemic cells abolishes SOCE and significantly prolongs the survival of mice in a NOTCH1-dependent model of T-ALL. The survival advantage is unrelated to the leukemic cell burden but is associated with the SOCE-dependent ability of malignant T lymphoblasts to cause inflammation in leukemia-infiltrated organs. Mice with STIM1/STIM2-deficient T-ALL show a markedly reduced necroinflammatory response in leukemia-infiltrated organs and downregulation of signaling pathways previously linked to cancer-induced inflammation. Our study shows that leukemic T lymphoblasts cause inflammation of leukemia-infiltrated organs that is dependent on SOCE. KW - T cell acute lymphoblastic leukemia KW - T-ALL KW - Notch1 KW - STIM1 KW - STIM2 KW - calcium KW - Ca2+ KW - CRAC KW - channel KW - inflammation KW - interferon KW - anemia KW - macrophages Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-227259 VL - 24 IS - 11 ER - TY - JOUR A1 - Herrmann, Johannes A1 - Muenstermann, Marcel A1 - Strobel, Lea A1 - Schubert-Unkmeir, Alexandra A1 - Woodruff, Trent M. A1 - Gray-Owen, Scott D. A1 - Klos, Andreas A1 - Johswich, Kay O. T1 - Complement C5a receptor 1 exacerbates the pathophysiology of N. meningitidis sepsis and is a potential target for disease treatment JF - mBio N2 - Sepsis caused by Neisseria meningitidis (meningococcus) is a rapidly progressing, life-threatening disease. Because its initial symptoms are rather unspecific, medical attention is often sought too late, i.e., when the systemic inflammatory response is already unleashed. This in turn limits the success of antibiotic treatment. The complement system is generally accepted as the most important innate immune determinant against invasive meningococcal disease since it protects the host through the bactericidal membrane attack complex. However, complement activation concomitantly liberates the C5a peptide, and it remains unclear whether this potent anaphylatoxin contributes to protection and/or drives the rapidly progressing immunopathogenesis associated with meningococcal disease. Here, we dissected the specific contribution of C5a receptor 1 (C5aR1), the canonical receptor for C5a, using a mouse model of meningococcal sepsis. Mice lacking C3 or C5 displayed susceptibility that was enhanced by >1,000-fold or 100-fold, respectively, consistent with the contribution of these components to protection. In clear contrast, C5ar1\(^{-/-}\) mice resisted invasive meningococcal infection and cleared N. meningitidis more rapidly than wild-type (WT) animals. This favorable outcome stemmed from an ameliorated inflammatory cytokine response to N. meningitidis in C5ar1\(^{-/-}\) mice in both in vivo and ex vivo whole-blood infections. In addition, inhibition of C5aR1 signaling without interference with the complement bactericidal activity reduced the inflammatory response also in human whole blood. Enticingly, pharmacologic C5aR1 blockade enhanced mouse survival and lowered meningococcal burden even when the treatment was administered after sepsis induction. Together, our findings demonstrate that C5aR1 drives the pathophysiology associated with meningococcal sepsis and provides a promising target for adjunctive therapy. Importance: The devastating consequences of N. meningitidis sepsis arise due to the rapidly arising and self-propagating inflammatory response that mobilizes antibacterial defenses but also drives the immunopathology associated with meningococcemia. The complement cascade provides innate broad-spectrum protection against infection by directly damaging the envelope of pathogenic microbes through the membrane attack complex and triggers an inflammatory response via the C5a peptide and its receptor C5aR1 aimed at mobilizing cellular effectors of immunity. Here, we consider the potential of separating the bactericidal activities of the complement cascade from its immune activating function to improve outcome of N. meningitidis sepsis. Our findings demonstrate that the specific genetic or pharmacological disruption of C5aR1 rapidly ameliorates disease by suppressing the pathogenic inflammatory response and, surprisingly, allows faster clearance of the bacterial infection. This outcome provides a clear demonstration of the therapeutic benefit of the use of C5aR1-specific inhibitors to improve the outcome of invasive meningococcal disease. KW - C5aR1 KW - whole-blood model KW - Neisseria meningitidis KW - anaphylatoxins KW - complement system KW - inflammation KW - invasive disease KW - mouse model KW - neutrophils KW - sepsis Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-175792 VL - 9 IS - 1 ER - TY - JOUR A1 - Ungern-Sternberg, Saskia N. I. von A1 - Zernecke, Alma A1 - Seizer, Peter T1 - Extracellular matrix metalloproteinase inducer EMMPRIN (CD147) in cardiovascular disease JF - International Journal of Molecular Sciences N2 - The receptor EMMPRIN is involved in the development and progression of cardiovascular diseases and in the pathogenesis of myocardial infarction. There are several binding partners of EMMPRIN mediating the effects of EMMPRIN in cardiovascular diseases. EMMPRIN interaction with most binding partners leads to disease progression by mediating cytokine or chemokine release, the activation of platelets and monocytes, as well as the formation of monocyte-platelet aggregates (MPAs). EMMPRIN is also involved in atherosclerosis by mediating the infiltration of pro-inflammatory cells. There is also evidence that EMMPRIN controls energy metabolism of cells and that EMMPRIN binding partners modulate intracellular glycosylation and trafficking of EMMPRIN towards the cell membrane. In this review, we systematically discuss these multifaceted roles of EMMPRIN and its interaction partners, such as Cyclophilins, in cardiovascular disease. KW - cardiovascular disease KW - immunoglobulin superfamily KW - inflammation KW - platelets KW - monocyte-platelet aggregates Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-285014 SN - 1422-0067 VL - 19 IS - 2 ER - TY - JOUR A1 - Groh, Janos A1 - Hörner, Michaela A1 - Martini, Rudolf T1 - Teriflunomide attenuates neuroinflammation-related neural damage in mice carrying human PLP1 mutations JF - Journal of Neuroinflammation N2 - Background: Genetically caused neurological disorders of the central nervous system (CNS) are mostly characterized by poor or even fatal clinical outcome and few or no causative treatments are available. Often, these disorders are associated with low-grade, disease-promoting inflammation, another feature shared by progressive forms of multiple sclerosis (PMS). We previously generated two mouse lines carrying distinct mutations in the oligodendrocytic PLP1 gene that have initially been identified in patients diagnosed with MS. These mutations cause a loss of PLP function leading to a histopathological and clinical phenotype common to both PMS and genetic CNS disorders, like hereditary spastic paraplegias. Importantly, neuroinflammation promotes disease progression in these models, suggesting that pharmacological modulation of inflammation might ameliorate disease outcome. Methods: We applied teriflunomide, an approved medication for relapsing-remitting MS targeting activated T-lymphocytes, in the drinking water (10 mg/kg body weight/day). Experimental long-term treatment of PLP mutant mice was non-invasively monitored by longitudinal optical coherence tomography and by rotarod analysis. Immunomodulatory effects were subsequently analyzed by flow cytometry and immunohistochemistry and treatment effects regarding neural damage, and neurodegeneration were assessed by histology and immunohistochemistry. Results: Preventive treatment with teriflunomide attenuated the increase in number of CD8+ cytotoxic effector T cells and fostered the proliferation of CD8+ CD122+ PD-1+ regulatory T cells in the CNS. This led to an amelioration of axonopathic features and neuron loss in the retinotectal system, also reflected by reduced thinning of the innermost retinal composite layer in longitudinal studies and ameliorated clinical outcome upon preventive long-term treatment. Treatment of immune-incompetent PLP mutants did not provide evidence for a direct, neuroprotective effect of the medication. When treatment was terminated, no rebound of neuroinflammation occurred and histopathological improvement was preserved for at least 75 days without treatment. After disease onset, teriflunomide halted ongoing axonal perturbation and enabled a recovery of dendritic arborization by surviving ganglion cells. However, neither neuron loss nor clinical features were ameliorated, likely due to already advanced neurodegeneration before treatment onset. Conclusions: We identify teriflunomide as a possible medication not only for PMS but also for inflammation-related genetic diseases of the nervous system for which causal treatment options are presently lacking. KW - axonal degeneration KW - inflammation KW - proteolipid protein KW - T-lymphocytes KW - teriflunomide Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-176524 VL - 15 IS - 194 ER - TY - THES A1 - Karl, Franziska T1 - The role of miR-21 in the pathophysiology of neuropathic pain using the model of B7-H1 knockout mice T1 - Die Rolle von miR-21 in der Pathophysiologie von neuropathischem Schmerz am Model der B7-H1 defizienten Maus N2 - The impact of microRNA (miRNA) as key players in the regulation of immune and neuronal gene expression and their role as master switches in the pathophysiology of neuropathic pain is increasingly recognized. miR-21 is a promising candidate that could be linked to the immune and the nociceptive system. To further investigate the pathophysiological role of miR-21 in neuropathic pain, we assesed mice deficient of B7 homolog 1 (B7-H1 ko), a protein with suppressive effect on inflammatory responses. B7-H1 ko mice and wildtype littermates (WT) of three different age-groups, young (8 weeks), middle-aged (6 months), and old (12 months) received a spared nerve injury (SNI). Thermal withdrawal latencies and mechanical withdrawal thresholds were determined. Further, we investigated anxiety-, depression-like and cognitive behavior. Quantitative real time PCR was used to determine miR-21 relative expression in peripheral nerves, dorsal root ganglia and white blood cells (WBC) at distinct time points after SNI. Naïve B7-H1 ko mice showed mechanical hyposensitivity with increasing age. Young and middle-aged B7-H1 ko mice displayed lower mechanical withdrawal thresholds compared to WT mice. From day three after SNI both genotypes developed mechanical and heat hypersensitivity, without intergroup differences. As supported by the results of three behavioral tests, no relevant differences were found for anxiety-like behavior after SNI in B7-H1 ko and WT mice. Also, there was no indication of depression-like behavior after SNI or any effect of SNI on cognition in both genotypes. The injured nerves of B7-H1 ko and WT mice showed higher miR-21 expression and invasion of macrophages and T cells 7 days after SNI without intergroup differences. Perineurial miR-21 inhibitor injection reversed SNI-induced mechanical and heat hypersensitivity in old B7-H1 ko and WT mice. This study reveals that reduced mechanical thresholds and heat withdrawal latencies are associated with miR-21 induction in the tibial and common peroneal nerve after SNI, which can be reversed by perineurial injection of a miR-21 inhibitor. Contrary to expectations, miR-21 expression levels were not higher in B7-H1 ko compared to WT mice. Thus, the B7-H1 ko mouse may be of minor importance for the study of miR-21 related pain. However, these results spot the contribution of miR-21 in the pathophysiology of neuropathic pain and emphasize the crucial role of miRNA in the regulation of neuronal and immune circuits that contribute to neuropathic pain. N2 - Die Beteiligung von microRNA (miRNA) an der Genregulation immunologischer und neuronaler Prozesse und deren Rolle als Schlüsselelement in der Pathophysiologie von neuropathischem Schmerz gewinnt zunehmend an Bedeutung. miR-21 ist ein vielversprechender Kandidat, der sowohl das Immunsystem, als auch das nozizeptive System beeinflusst. Um die pathophysiologische Rolle von miR-21 bei neuropathischem Schmerz besser zu verstehen wurden Mäuse mit B7 homolog 1 Defizienz (B7-H1 ko), einem immunsupprimierendem Protein, untersucht. Eine frühere Studie zeigte eine Hochregulierung von miR-21 in murinen Lymphozyten. Junge (8 Wochen), mittelalte (6 Monate) und alte (12 Monate) B7-H1 ko Mäuse und Wildtypwurfgeschwister (WT) erhielten eine spared nerve injury (SNI) als neuropathischem Schmerzmodell. Es wurden thermische Rückzugslatenzen und mechanische Rückzugsschwellen bestimmt. Des weiteren wurde sowohl das Angstverhalten, das depressive Verhalten, als auch das kognitive Verhalten untersucht. Um die relative Expression von miR-21 in den peripheren Nerven, den Spinalganglien und in den weißen Blutzellen zu verschiedenen Zeitpunkten zu bestimmen, wurde die quantitative real time PCR angewandt. Naive B7-H1 ko Mäuse zeigten mit zunehmendem Alter eine mechanische Hyposensitivität. Bereits 3 Tage nach SNI entwickelten beide Genotypen eine Überempfindlichkeit gegenüber Hitze und mechanischer Stimulation. In drei durchgeführten Verhaltenstests konnten keine relevanten Unterschiede im Angstverhalten nach SNI von B7-H1 ko und WT Mäusen festgestellt werden. Bei beiden Genotypen gab es weder Hinweise auf depressives Verhalten nach SNI, noch wurde das kognitive Verhalten durch SNI beeinträchtigt. Die verletzen Nerven der B7-H1 ko und WT Mäuse zeigten 7 Tage nach SNI eine höhere miR-21 Expression und eine Invasion durch Makrophagen und T-Zellen ohne Gruppenunterschiede. Die perineurale Injektion eines miR-21 Inhibitors konnte die durch SNI induzierte mechanische und thermische Hypersensitivität lindern. Diese Studie zeigt, dass der Anstieg von miR-21 im N. tibialis und N. peroneus communis mit reduzierten Rückzugsschwellen gegen mechanische Reize und verkürzten Wegzugslatenzen bei Hitzestimulation einhergeht, welche durch perineurale Injektion eines miR-21 Inhibitors verringert werden können. Entgegen der Erwartungen zeigten B7-H1 ko Mäuse im Vergleich zu WT Mäusen keine erhöhte miR-21 Expression und sind daher möglicherweise von geringer Bedeutung für die Untersuchung von miR-21 assoziiertem Schmerz. Jedoch bekräftigen diese Ergebnisse eine Beteiligung von miR-21 an der Pathophysiologie von neuropathischem Schmerz und bestätigen die wichtige Rolle von miRNA bei der Regulation von neuronalen und immunologischen Prozessen, die zu neuropathischem Schmerz beitragen. KW - neuropathic pain KW - inflammation KW - B7-H1 KW - immune system KW - neuropathic pain KW - miRNA KW - miR-21 Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-156004 ER - TY - JOUR A1 - Hofmann, Sigrun Ruth A1 - Böttger, Fanny A1 - Range, Ursula A1 - Lück, Christian A1 - Morbach, Henner A1 - Girschick, Hermann Joseph A1 - Suttorp, Meinolf A1 - Hedrich, Christian Michael T1 - Serum interleukin-6 and CCL11/eotaxin may be suitable biomarkers for the diagnosis of chronic nonbacterial osteomyelitis JF - Frontiers in Pediatrics N2 - Objectives: Chronic recurrent multifocal osteomyelitis (CRMO), the most severe form of chronic nonbacterial osteomyelitis (CNO), is an autoinflammatory bone disorder. In the absence of diagnostic criteria or biomarkers, CNO/CRMO remains a diagnosis of exclusion. The aim of this study was to identify biomarkers for diagnosing multifocal disease (CRMO). Study design: Sera from 71 pediatric CRMO patients, 11 patients with osteoarticular infections, 62 patients with juvenile idiopathic arthritis (JIA), 7 patients with para-infectious or reactive arthritis, and 43 patients with acute leukemia or lymphoma, as well as 59 healthy individuals were collected. Multiplex analysis of 18 inflammation- and/or bone remodeling-associated serum proteins was performed. Statistical analysis included univariate ANOVA, discriminant analysis, univariate receiver operating characteristic (ROC) analysis, and logistic regression analyses. Results: For 14 of 18 blood serum proteins, significant differences were determined between CRMO patients, at least one alternative diagnosis, or healthy controls. Multi-component discriminant analysis delivered five biomarkers (IL-6, CCL11/eotaxin, CCL5/RANTES, collagen Iα, sIL-2R) for the diagnosis of CRMO. ROC analysis allowed further reduction to a core set of 2 biomarkers (CCL11/eotaxin, IL-6) that are sufficient to discern between CRMO, healthy controls, and alternative diagnoses. Conclusion: Serum biomarkers CCL11/eotaxin and IL-6 differentiate between patients with CRMO, healthy controls, and alternative diagnoses (leukemia and lymphoma, osteoarticular infections, para-infectious arthritis, and JIA). Easily accessible biomarkers may aid in diagnosing CRMO. Further studies testing biomarkers in larger unrelated cohorts are warranted. KW - medicine KW - chronic nonbacterial osteomyelitis KW - chronic recurrent multifocal osteomyelitis KW - inflammation KW - biomarker KW - autoinflammation KW - diagnosis Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-172744 VL - 5 ER - TY - JOUR A1 - Koeniger, Tobias A1 - Kuerten, Stefanie T1 - Splitting the "unsplittable": Dissecting resident and infiltrating macrophages in experimental autoimmune encephalomyelitis JF - International Journal of Molecular Sciences N2 - Macrophages predominate the inflammatory landscape within multiple sclerosis (MS) lesions, not only regarding cellularity but also with respect to the diverse functions this cell fraction provides during disease progression and remission. Researchers have been well aware of the fact that the macrophage pool during central nervous system (CNS) autoimmunity consists of a mixture of myeloid cells. Yet, separating these populations to define their unique contribution to disease pathology has long been challenging due to their similar marker expression. Sophisticated lineage tracing approaches as well as comprehensive transcriptome analysis have elevated our insight into macrophage biology to a new level enabling scientists to dissect the roles of resident (microglia and non-parenchymal macrophages) and infiltrating macrophages with unprecedented precision. To do so in an accurate way, researchers have to know their toolbox, which has been filled with diverse, discriminating approaches from decades of studying neuroinflammation in animal models. Every method has its own strengths and weaknesses, which will be addressed in this review. The focus will be on tools to manipulate and/or identify different macrophage subgroups within the injured murine CNS. KW - CNS KW - distinction KW - experimental autoimmune encephalomyelitis KW - inflammation KW - macrophages KW - markers KW - microglia KW - monocytes Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-285067 SN - 1422-0067 VL - 18 IS - 10 ER - TY - JOUR A1 - Schmid, Tobias A1 - Falter, Lena A1 - Weber, Sabine A1 - Müller, Nils A1 - Molitor, Konstantin A1 - Zeller, David A1 - Weber-Steffens, Dorothea A1 - Hehlgans, Thomas A1 - Wajant, Harald A1 - Mostböck, Sven A1 - Männel, Daniela N. T1 - Chronic inflammation increases the sensitivity of mouse Treg for TNFR2 costimulation JF - Frontiers in Immunology N2 - TNF receptor type 2 (TNFR2) has gained attention as a costimulatory receptor for T cells and as critical factor for the development of regulatory T cells (Treg) and myeloid suppressor cells. Using the TNFR2-specific agonist TNCscTNF80, direct effects of TNFR2 activation on myeloid cells and T cells were investigated in mice. \(In\) \(vitro\), TNCscTNF80 induced T cell proliferation in a costimulatory fashion, and also supported \(in\) \(vitro\) expansion of Treg cells. In addition, activation of TNFR2 retarded differentiation of bone marrow-derived immature myeloid cells in culture and reduced their suppressor function. \(In\) \(vivo\) application of TNCscTNF80-induced mild myelopoiesis in naïve mice without affecting the immune cell composition. Already a single application expanded Treg cells and improved suppression of CD4 T cells in mice with chronic inflammation. By contrast, multiple applications of the TNFR2 agonist were required to expand Treg cells in naïve mice. Improved suppression of T cell proliferation depended on expression of TNFR2 by T cells in mice repeatedly treated with TNCscTNF80, without a major contribution of TNFR2 on myeloid cells. Thus, TNFR2 activation on T cells in naïve mice can lead to immune suppression \(in\) \(vivo\). These findings support the important role of TNFR2 for Treg cells in immune regulation. KW - molecular medicine KW - inflammation KW - immune regulation KW - costimulation KW - MDSC KW - TNFR2 KW - regulatory T cell Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-173259 VL - 8 ER - TY - JOUR A1 - Oehler, Beatrice A1 - Mohammadi, Milad A1 - Perpina Viciano, Cristina A1 - Hackel, Dagmar A1 - Hoffmann, Carsten A1 - Brack, Alexander A1 - Rittner, Heike L. T1 - Peripheral interaction of Resolvin D1 and E1 with opioid receptor antagonists for antinociception in inflammatory pain in rats JF - Frontiers in Molecular Neuroscience N2 - Antinociceptive pathways are activated in the periphery in inflammatory pain, for instance resolvins and opioid peptides. Resolvins are biosynthesized from omega-3 polyunsaturated fatty acids such as eicosapentaenoic acid and docosahexaenoic acid. Resolvin D1 (RvD1) and resolvin E1 (RvE1) initiate the resolution of inflammation and control of hypersensitivity via induction of anti-inflammatory signaling cascades. RvD1 binds to lipoxin A4/annexin-A1 receptor/formyl-peptide receptor 2 (ALX/FPR2), RvE1 to chemerin receptor 23 (ChemR23). Antinociception of RvD1 is mediated by interaction with transient receptor potential channels ankyrin 1 (TRPA1). Endogenous opioid peptides are synthesized and released from leukocytes in the tissue and bind to opioid receptors on nociceptor terminals. Here, we further explored peripheral mechanisms of RvD1 and chemerin (Chem), the ligand of ChemR23, in complete Freund’s adjuvant (CFA)-induced hindpaw inflammation in male Wistar rats. RvD1 and Chem ameliorated CFA-induced hypersensitivity in early and late inflammatory phases. This was prevented by peripheral blockade of the μ-opioid peptide receptor (MOR) using low dose local naloxone or by local injection of anti-β-endorphin and anti-met-enkephalin (anti-ENK) antibodies. Naloxone also hindered antinociception by the TRPA1 inhibitor HC-030031. RvD1 did not stimulate the release of β-endorphin from macrophages and neutrophils, nor did RvD1 itself activate G-proteins coupled MOR or initiate β-arrestin recruitment to the membrane. TRPA1 blockade by HC-030031 in inflammation in vivo as well as inhibition of the TRPA1-mediated calcium influx in dorsal root ganglia neurons in vitro was hampered by naloxone. Peripheral application of naloxone alone in vivo already lowered mechanical nociceptive thresholds. Therefore, either a perturbation of the balance of endogenous pro- and antinociceptive mechanisms in early and late inflammation, or an interaction of TRPA1 and opioid receptors weaken the antinociceptive potency of RvD1 and TRPA1 blockers. KW - transient receptor potential channels KW - pain behavior KW - resolvin KW - opioid receptors KW - opioid peptides KW - inflammation KW - animals Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-158642 VL - 10 IS - 242 ER - TY - JOUR A1 - Schuhmann, Michael K. A1 - Fluri, Felix T1 - Effects of fullerenols on mouse brain microvascular endothelial cells JF - International Journal of Molecular Sciences N2 - Fullerenols, water-soluble C60-fullerene derivatives, have been shown to exert neuroprotective effects in vitro and in vivo, most likely due to their capability to scavenge free radicals. However, little is known about the effects of fullerenols on the blood–brain barrier (BBB), especially on cerebral endothelial cells under inflammatory conditions. Here, we investigated whether the treatment of primary mouse brain microvascular endothelial cells with fullerenols impacts basal and inflammatory blood–brain barrier (BBB) properties in vitro. While fullerenols (1, 10, and 100 µg/mL) did not change transendothelial electrical resistance under basal and inflammatory conditions, 100 µg/mL of fullerenol significantly reduced erk1/2 activation and resulted in an activation of NFκB in an inflammatory milieu. Our findings suggest that fullerenols might counteract oxidative stress via the erk1/2 and NFκB pathways, and thus are able to protect microvascular endothelial cells under inflammatory conditions. KW - mouse brain microvascular endothelial cell cultur KW - adhesion molecules KW - fullerenes KW - blood-brain barrier KW - inflammation KW - tight junctions Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-158072 SN - 1422-0067 VL - 18 IS - 8 ER - TY - JOUR A1 - Glaser, Kirsten A1 - Silwedel, Christine A1 - Fehrholz, Markus A1 - Waaga-Gasser, Ana M. A1 - Henrich, Birgit A1 - Claus, Heike A1 - Speer, Christian P. T1 - Ureaplasma Species Differentially Modulate Pro- and Anti-Inflammatory Cytokine Responses in Newborn and Adult Human Monocytes Pushing the State Toward Pro-Inflammation JF - Frontiers in Cellular and Infection Microbiology N2 - Background: Ureaplasma species have been associated with chorioamnionitis and preterm birth and have been implicated in the pathogenesis of neonatal short and long-term morbidity. However, being mostly commensal bacteria, controversy remains on the pro-inflammatory capacity of Ureaplasma. Discussions are ongoing on the incidence and impact of prenatal, perinatal, and postnatal infection. The present study addressed the impact of Ureaplasma isolates on monocyte-driven inflammation. Methods: Cord blood monocytes of term neonates and adult monocytes, either native or LPS-primed, were cultured with Ureaplasma urealyticum (U. urealyticum) serovar 8 (Uu8) and Ureaplasma parvum serovar 3 (Up3). Using qRT-PCR, cytokine flow cytometry, and multi-analyte immunoassay, we assessed mRNA and protein expression of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-8, IL-12p40, IL-10, and IL-1 receptor antagonist (IL-1ra) as well as Toll-like receptor (TLR) 2 and TLR4. Results: Uu8 and Up3 induced mRNA expression and protein release of TNF-α, IL-1β and IL-8 in term neonatal and adult monocytes (p < 0.01 and p < 0.05). Intracellular protein expression of TNF-α, IL-1β and IL-8 in Ureaplasma-stimulated cells paralleled those results. Ureaplasma-induced cytokine levels did not significantly differ from LPS-mediated levels except for lower intracellular IL-1β in adult monocytes (Uu8: p < 0.05). Remarkably, ureaplasmas did not induce IL-12p40 response and promoted lower amounts of anti-inflammatory IL-10 and IL-1ra than LPS, provoking a cytokine imbalance more in favor of pro-inflammation (IL-1β/IL-10, IL-8/IL-10 and IL-8/IL-1ra: p < 0.01, vs. LPS). In contrast to LPS, both isolates induced TLR2 mRNA in neonatal and adult cells (p < 0.001 and p < 0.05) and suppressed TLR4 mRNA in adult monocytes (p < 0.05). Upon co-stimulation, Uu8 and Up3 inhibited LPS-induced intracellular IL-1β (p < 0.001 and p < 0.05) and IL-8 in adult monocytes (p < 0.01), while LPS-induced neonatal cytokines were maintained or aggravated (p < 0.05). Conclusion: Our data demonstrate a considerable pro-inflammatory capacity of Ureaplasma isolates in human monocytes. Stimulating pro-inflammatory cytokine responses while hardly inducing immunomodulatory and anti-inflammatory cytokines, ureaplasmas might push monocyte immune responses toward pro-inflammation. Inhibition of LPS-induced cytokines in adult monocytes in contrast to sustained inflammation in term neonatal monocytes indicates a differential modulation of host immune responses to a second stimulus. Modification of TLR2 and TLR4 expression may shape host susceptibility to inflammation. KW - Ureaplasma KW - infection KW - inflammation KW - immunomodulation KW - chorioamnionitis KW - neonatal morbidity KW - monocytes KW - cord blood Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-169958 VL - 7 IS - 484 ER - TY - JOUR A1 - Jannasch, Maren A1 - Gaetzner, Sabine A1 - Weigel, Tobias A1 - Walles, Heike A1 - Schmitz, Tobias A1 - Hansmann, Jan T1 - A comparative multi-parametric in vitro model identifies the power of test conditions to predict the fibrotic tendency of a biomaterial JF - Scientific Reports N2 - Despite growing effort to advance materials towards a low fibrotic progression, all implants elicit adverse tissue responses. Pre-clinical biomaterial assessment relies on animals testing, which can be complemented by in vitro tests to address the Russell and Burch’s 3R aspect of reducing animal burden. However, a poor correlation between in vitro and in vivo biomaterial assessments confirms a need for suitable in vitro biomaterial tests. The aim of the study was to identify a test setting, which is predictive and might be time- and cost-efficient. We demonstrated how sensitive in vitro biomaterial assessment based on human primary macrophages depends on test conditions. Moreover, possible clinical scenarios such as lipopolysaccharide contamination, contact to autologous blood plasma, and presence of IL-4 in an immune niche influence the outcome of a biomaterial ranking. Nevertheless, by using glass, titanium, polytetrafluorethylene, silicone, and polyethylene representing a specific material-induced fibrotic response and by comparison to literature data, we were able to identify a test condition that provides a high correlation to state-of-the-art in vivo studies. Most important, biomaterial ranking obtained under native plasma test conditions showed a high predictive accuracy compared to in vivo assessments, strengthening a biomimetic three-dimensional in vitro test platform. KW - inflammation KW - experimental models of disease KW - biomaterial tests KW - in vitro Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-170908 VL - 7 IS - 1689 ER - TY - JOUR A1 - Brodehl, Andreas A1 - Belke, Darrell D. A1 - Garnett, Lauren A1 - Martens, Kristina A1 - Abdelfatah, Nelly A1 - Rodriguez, Marcela A1 - Diao, Catherine A1 - Chen, Yong-Xiang A1 - Gordon, Paul M. K. A1 - Nygren, Anders A1 - Gerull, Brenda T1 - Transgenic mice overexpressing desmocollin-2 (DSC2) develop cardiomyopathy associated with myocardial inflammation and fibrotic remodeling JF - PLoS ONE N2 - Background Arrhythmogenic cardiomyopathy is an inherited heart muscle disorder leading to ventricular arrhythmias and heart failure, mainly as a result of mutations in cardiac desmosomal genes. Desmosomes are cell-cell junctions mediating adhesion of cardiomyocytes; however, the molecular and cellular mechanisms underlying the disease remain widely unknown. Desmocollin-2 is a desmosomal cadherin serving as an anchor molecule required to reconstitute homeostatic intercellular adhesion with desmoglein-2. Cardiac specific lack of desmoglein-2 leads to severe cardiomyopathy, whereas overexpression does not. In contrast, the corresponding data for desmocollin-2 are incomplete, in particular from the view of protein overexpression. Therefore, we developed a mouse model overexpressing desmocollin-2 to determine its potential contribution to cardiomyopathy and intercellular adhesion pathology. Methods and results We generated transgenic mice overexpressing DSC2 in cardiac myocytes. Transgenic mice developed a severe cardiac dysfunction over 5 to 13 weeks as indicated by 2D-echocardiography measurements. Corresponding histology and immunohistochemistry demonstrated fibrosis, necrosis and calcification which were mainly localized in patches near the epi- and endocardium of both ventricles. Expressions of endogenous desmosomal proteins were markedly reduced in fibrotic areas but appear to be unchanged in non-fibrotic areas. Furthermore, gene expression data indicate an early up-regulation of inflammatory and fibrotic remodeling pathways between 2 to 3.5 weeks of age. Conclusion Cardiac specific overexpression of desmocollin-2 induces necrosis, acute inflammation and patchy cardiac fibrotic remodeling leading to fulminant biventricular cardiomyopathy. KW - heart KW - mouse models KW - gene expression KW - fibrosis KW - inflammation KW - gene expression KW - genetically modified animals KW - cardiomyopathies KW - hyperexpression techniques Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-171084 VL - 12 IS - 3 ER - TY - JOUR A1 - Fehrholz, Markus A1 - Glaser, Kirsten A1 - Seidenspinner, Silvia A1 - Ottensmeier, Barbara A1 - Curstedt, Tore A1 - Speer, Christian P. A1 - Kunzmann, Steffen T1 - Impact of the New Generation Reconstituted Surfactant CHF5633 on Human CD4\(^+\) Lymphocytes JF - PLoS One N2 - Background Natural surfactant preparations, commonly isolated from porcine or bovine lungs, are used to treat respiratory distress syndrome in preterm infants. Besides biophysical effectiveness, several studies have documented additional immunomodulatory properties. Within the near future, synthetic surfactant preparations may be a promising alternative. CHF5633 is a new generation reconstituted synthetic surfactant preparation with defined composition, containing dipalmitoyl-phosphatidylcholine, palmitoyl-oleoyl-phosphatidylglycerol and synthetic analogs of surfactant protein (SP-) B and SP-C. While its biophysical effectiveness has been demonstrated in vitro and in vivo, possible immunomodulatory abilities are currently unknown. Aim The aim of the current study was to define a potential impact of CHF5633 and its single components on pro- and anti-inflammatory cytokine responses in human CD4\(^+\) lymphocytes. Methods Purified human CD4\(^+\) T cells were activated using anti CD3/CD28 antibodies and exposed to CHF5633, its components, or to the well-known animal-derived surfactant Poractant alfa (Curosurf®). Proliferative response and cell viability were assessed using flow cytometry and a methylthiazolyldiphenyltetrazolium bromide colorimetric assay. The mRNA expression of IFNγ, IL-2, IL-17A, IL-22, IL-4, and IL-10 was measured by quantitative PCR, while intracellular protein expression was assessed by means of flow cytometry. Results Neither CHF5633 nor any of its phospholipid components with or without SP-B or SP-C analogs had any influence on proliferative ability and viability of CD4\(^+\) lymphocytes under the given conditions. IFNγ, IL-2, IL-17A, IL-22, IL-4, and IL-10 mRNA as well as IFNγ, IL-2, IL-4 and IL-10 protein levels were unaffected in both non-activated and activated CD4+ lymphocytes after exposure to CHF5633 or its constituents compared to non-exposed controls. However, in comparison to Curosurf®, expression levels of anti-inflammatory IL-4 and IL-10 mRNA were significantly increased in CHF5633 exposed CD4\(^+\) lymphocytes. Conclusion For the first time, the immunomodulatory capacity of CHF5633 on CD4\(^+\) lymphocytes was evaluated. CHF5633 did not show any cytotoxicity on CD4\(^+\) cells. Moreover, our in vitro data indicate that CHF5633 does not exert unintended pro-inflammatory effects on non-activated and activated CD4+ T cells. As far as anti-inflammatory cytokines are concerned, it might lack an overall reductive ability in comparison to animal-derived surfactants, potentially leaving pro- and anti-inflammatory cytokine response in balance. KW - lymphocytes KW - surfactants KW - flow cytometry KW - monocytes KW - RNA isolation KW - T cells KW - cytokines KW - inflammation Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-146419 VL - 11 IS - 4 ER - TY - JOUR A1 - Suliman, Salwa A1 - Sun, Yang A1 - Pedersen, Torbjorn O. A1 - Xue, Ying A1 - Nickel, Joachim A1 - Waag, Thilo A1 - Finne-Wistrand, Anna A1 - Steinmüller-Nethl, Doris A1 - Krueger, Anke A1 - Costea, Daniela E. A1 - Mustafa, Kamal T1 - In vivo host response and degradation of copolymer scaffolds functionalized with nanodiamonds and bone morphogenetic protein 2 JF - Advanced Healthcare Materials N2 - The aim is to evaluate the effect of modifying poly[(L-lactide)-co-(epsilon-caprolactone)] scaffolds (PLCL) with nanodiamonds (nDP) or with nDP+physisorbed BMP-2 (nDP+BMP-2) on in vivo host tissue response and degradation. The scaffolds are implanted subcutaneously in Balb/c mice and retrieved after 1, 8, and 27 weeks. Molecular weight analysis shows that modified scaffolds degrade faster than the unmodified. Gene analysis at week 1 shows highest expression of proinflammatory markers around nDP scaffolds; although the presence of inflammatory cells and foreign body giant cells is more prominent around the PLCL. Tissue regeneration markers are highly expressed in the nDP+BMP-2 scaffolds at week 8. A fibrous capsule is detectable by week 8, thinnest around nDP scaffolds and at week 27 thickest around PLCL scaffolds. mRNA levels of ALP, COL1 alpha 2, and ANGPT1 are signifi cantly upregulating in the nDP+BMP-2 scaffolds at week 1 with ectopic bone seen at week 8. Even when almost 90% of the scaffold is degraded at week 27, nDP are observable at implantation areas without adverse effects. In conclusion, modifying PLCL scaffolds with nDP does not aggravate the host response and physisorbed BMP-2 delivery attenuates infl ammation while lowering the dose of BMP-2 to a relatively safe and economical level. KW - inflammatory response KW - biocompatibility KW - BMP-2 delivery KW - inflammation KW - tissue engineering Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-189764 VL - 5 IS - 6 ER - TY - JOUR A1 - Rosenbaum, Corinna A1 - Schick, Martin Alexander A1 - Wollborn, Jakob A1 - Heider, Andreas A1 - Scholz, Claus-Jürgen A1 - Cecil, Alexander A1 - Niesler, Beate A1 - Hirrlinger, Johannes A1 - Walles, Heike A1 - Metzger, Marco T1 - Activation of Myenteric Glia during Acute Inflammation In Vitro and In Vivo JF - PLoS One N2 - Background Enteric glial cells (EGCs) are the main constituent of the enteric nervous system and share similarities with astrocytes from the central nervous system including their reactivity to an inflammatory microenvironment. Previous studies on EGC pathophysiology have specifically focused on mucosal glia activation and its contribution to mucosal inflammatory processes observed in the gut of inflammatory bowel disease (IBD) patients. In contrast knowledge is scarce on intestinal inflammation not locally restricted to the mucosa but systemically affecting the intestine and its effect on the overall EGC network. Methods and Results In this study, we analyzed the biological effects of a systemic LPS-induced hyperinflammatory insult on overall EGCs in a rat model in vivo, mimicking the clinical situation of systemic inflammation response syndrome (SIRS). Tissues from small and large intestine were removed 4 hours after systemic LPS-injection and analyzed on transcript and protein level. Laser capture microdissection was performed to study plexus-specific gene expression alterations. Upon systemic LPS-injection in vivo we observed a rapid and dramatic activation of Glial Fibrillary Acidic Protein (GFAP)-expressing glia on mRNA level, locally restricted to the myenteric plexus. To study the specific role of the GFAP subpopulation, we established flow cytometry-purified primary glial cell cultures from GFAP promotor-driven EGFP reporter mice. After LPS stimulation, we analyzed cytokine secretion and global gene expression profiles, which were finally implemented in a bioinformatic comparative transcriptome analysis. Enriched GFAP+ glial cells cultured as gliospheres secreted increased levels of prominent inflammatory cytokines upon LPS stimulation. Additionally, a shift in myenteric glial gene expression profile was induced that predominantly affected genes associated with immune response. Conclusion and Significance Our findings identify the myenteric GFAP-expressing glial subpopulation as particularly susceptible and responsive to acute systemic inflammation of the gut wall and complement knowledge on glial involvement in mucosal inflammation of the intestine. KW - gene expression KW - gastrointestinal tract KW - inflammatory bowel disease KW - central nervous system KW - systemic inflammatory response syndrome KW - inflammation KW - astrocytes KW - cytokines Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-146544 VL - 11 IS - 3 ER - TY - JOUR A1 - Alrefai, Hani A1 - Muhammad, Khalid A1 - Rudolf, Ronald A1 - Pham, Duong Anh Thuy A1 - Klein-Hessling, Stefan A1 - Patra, Amiya K. A1 - Avots, Andris A1 - Bukur, Valesca A1 - Sahin,, Ugur A1 - Tenzer, Stefan A1 - Goebeler, Matthias A1 - Kerstan, Andreas A1 - Serfling, Edgar T1 - NFATc1 supports imiquimod-induced skin inflammation by suppressing IL-10 synthesis in B cells JF - Nature Communications N2 - Epicutaneous application of Aldara cream containing the TLR7 agonist imiquimod (IMQ) to mice induces skin inflammation that exhibits many aspects of psoriasis, an inflammatory human skin disease. Here we show that mice depleted of B cells or bearing interleukin (IL)-10-deficient B cells show a fulminant inflammation upon IMQ exposure, whereas ablation of NFATc1 in B cells results in a suppression of Aldara-induced inflammation. In vitro, IMQ induces the proliferation and IL-10 expression by B cells that is blocked by BCR signals inducing NFATc1. By binding to HDAC1, a transcriptional repressor, and to an intronic site of the Il10 gene, NFATc1 suppresses IL-10 expression that dampens the production of tumour necrosis factor-α and IL-17 by T cells. These data indicate a close link between NFATc1 and IL-10 expression in B cells and suggest NFATc1 and, in particular, its inducible short isoform, NFATc1/αA, as a potential target to treat human psoriasis. KW - B cells KW - psoriasis KW - interleukins KW - inflammation Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-173053 VL - 7 ER - TY - JOUR A1 - Howangyin, Kiave-Yune A1 - Zlatanova, Ivana A1 - Pinto, Cristina A1 - Ngkelo, Anta A1 - Cochain, Clément A1 - Rouanet, Marie A1 - Vilar, José A1 - Lemitre, Mathilde A1 - Stockmann, Christian A1 - Fleischmann, Bernd K. A1 - Mallat, Ziad A1 - Silvestre, Jean-Sébastien T1 - Myeloid-epithelial-reproductive receptor tyrosine kinase and milk fat globule epidermal growth factor 8 coordinately improve remodeling after myocardial infarction via local delivery of vascular endothelial growth factor JF - Circulation N2 - Background: In infarcted heart, improper clearance of dying cells by activated neighboring phagocytes may precipitate the transition to heart failure. We analyzed the coordinated role of 2 major mediators of efferocytosis, the myeloid-epithelial-reproductive protein tyrosine kinase (Mertk) and the milk fat globule epidermal growth factor (Mfge8), in directing cardiac remodeling by skewing the inflammatory response after myocardial infarction. Methods and Results: We generated double-deficient mice for Mertk and Mfge8 (Mertk\(^{-/-}\)/Mfge8\(^{-/-}\)) and challenged them with acute coronary ligature. Compared with wild-type, Mertk-deficient (Mertk\(^{-/-}\)), or Mfge8-deficient (Mfge8\(^{-/-}\)) animals, Mertk\(^{-/-}\)/Mfge8\(^{-/-}\) mice displayed greater alteration in cardiac function and remodeling. Mertk and Mfge8 were expressed mainly by cardiac Ly6C\(^{High and Low}\) monocytes and macrophages. In parallel, Mertk\(^{-/-}\)/Mfge8\(^{-/-}\) bone marrow chimeras manifested increased accumulation of apoptotic cells, enhanced fibrotic area, and larger infarct size, as well as reduced angiogenesis. We found that the abrogation of efferocytosis affected neither the ability of circulating monocytes to infiltrate cardiac tissue nor the number of resident Ly6C\(^{High}\) and Ly6C\(^{Low}\) monocytes/macrophages populating the infarcted milieu. In contrast, combined Mertk and Mfge8 deficiency in Ly6C\(^{High}\)/Ly6C\(^{Low}\) monocytes/macrophages either obtained from in vitro differentiation of bone marrow cells or isolated from infarcted hearts altered their capacity of efferocytosis and subsequently blunted vascular endothelial growth factor A (VEGFA) release. Using LysMCre\(^+\)/VEGFA\(^{fl/fl}\) mice, we further identified an important role for myeloid-derived VEGFA in improving cardiac function and angiogenesis. Conclusions: After myocardial infarction, Mertk- and Mfge8-expressing monocyte/macrophages synergistically engage the clearance of injured cardiomyocytes, favoring the secretion of VEGFA to locally repair the dysfunctional heart. KW - inflammation KW - macrophages KW - myocardial infarction KW - myocarditis KW - neovascularization, physiologic Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-190755 VL - 133 IS - 9 ER - TY - JOUR A1 - Schuhmann, Michael K. A1 - Gunreben, Ignaz A1 - Kleinschnitz, Christoph A1 - Kraft, Peter T1 - Immunohistochemical Analysis of Cerebral Thrombi Retrieved by Mechanical Thrombectomy from Patients with Acute Ischemic Stroke JF - International Journal of Molecular Sciences N2 - Mechanical thrombectomy is a novel treatment option for patients with acute ischemic stroke (AIS). Only a few studies have previously suggested strategies to categorize retrieved clots according to their histologic composition. However, these reports did not analyze potential biomarkers that are of importance in stroke-related inflammation. We therefore histopathologically investigated 37 intracerebral thrombi mechanically retrieved from patients with AIS, and focused on the composition of immune cells and platelets. We also conducted correlation analyses of distinctive morphologic patterns (erythrocytic, serpentine, layered, red, white, mixed appearance) with clinical parameters. Most T cells and monocytes were detected in erythrocytic and red clots, in which the distribution of these cells was random. In contrast, von Willebrand factor (vWF)-positive areas co-localized with regions of fibrin and collagen. While clots with huge amounts of vWF seem to be associated with a high National Institute of Health Stroke Scale score at admission, histologic findings could not predict the clinical outcome at discharge. In summary, we provide the first histologic description of mechanically retrieved intracerebral thrombi regarding biomarkers relevant for inflammation in ischemic stroke. KW - thrombus formation KW - immune cells KW - lymphocytes KW - mechanical thrombectomy KW - ischemic stroke KW - inflammation Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-166206 VL - 17 IS - 3 ER - TY - JOUR A1 - Grimmig, Tanja A1 - Moench, Romana A1 - Kreckel, Jennifer A1 - Haack, Stephanie A1 - Rueckert, Felix A1 - Rehder, Roberta A1 - Tripathi, Sudipta A1 - Ribas, Carmen A1 - Chandraker, Anil A1 - Germer, Christoph T. A1 - Gasser, Martin A1 - Waaga-Gasser, Ana Maria T1 - Toll Like Receptor 2, 4, and 9 Signaling Promotes Autoregulative Tumor Cell Growth and VEGF/PDGF Expression in Human Pancreatic Cancer JF - International Journal of Molecular Sciences N2 - Toll like receptor (TLR) signaling has been suggested to play an important role in the inflammatory microenvironment of solid tumors and through this inflammation-mediated tumor growth. Here, we studied the role of tumor cells in their process of self-maintaining TLR expression independent of inflammatory cells and cytokine milieu for autoregulative tumor growth signaling in pancreatic cancer. We analyzed the expression of TLR2, -4, and -9 in primary human cancers and their impact on tumor growth via induced activation in several established pancreatic cancers. TLR-stimulated pancreatic cancer cells were specifically investigated for activated signaling pathways of VEGF/PDGF and anti-apoptotic Bcl-xL expression as well as tumor cell growth. The primary pancreatic cancers and cell lines expressed TLR2, -4, and -9. TLR-specific stimulation resulted in activated MAP-kinase signaling, most likely via autoregulative stimulation of demonstrated TLR-induced VEGF and PDGF expression. Moreover, TLR activation prompted the expression of Bcl-xL and has been demonstrated for the first time to induce tumor cell proliferation in pancreatic cancer. These findings strongly suggest that pancreatic cancer cells use specific Toll like receptor signaling to promote tumor cell proliferation and emphasize the particular role of TLR2, -4, and -9 in this autoregulative process of tumor cell activation and proliferation in pancreatic cancer. KW - tumor growth KW - TLR2 KW - TLR4 KW - TLR9 KW - pancreatic cancer KW - inflammation Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-165743 VL - 17 IS - 12 ER - TY - THES A1 - Panjwani, Priyadarshini T1 - Induction, Imaging, Histo-morphological and Molecular Characterization of Myocarditis in the Rat to Explore Novel Diagnostic Strategies for the Detection of Myocardial Inflammation T1 - Induktion, Bildgebung und, Histo-morphologische sowie Molekulare Charakterisierung der Myokarditis im Rattenmodell zur Entwicklung neuer diagnostischer Strategien zum Nachweis von Herzmuskelentzündungen N2 - Fulminant myocarditis is rare but a potentially life-threatening disease. Acute or mild myocarditis following acute ischemia is generally associated with a profound activation of the host’s immune system. On one hand this is mandatory to protect the host’s heart by fighting the invading agents (i.e., bacteria, viruses or other microbial agents) and/or to induce healing and repair processes in the damaged myocardium. On other hand, uncontrolled activation of the immune system may result in the generation of auto-reactive (not always beneficial) immune cells. Myocarditis or inflammatory cardiomyopathy is characterized by focal or diffuse infiltrates, myocyte necrosis and/or apoptosis and subsequent fibrotic replacement of the heart muscle. In humans, about 30% of the myocarditis-patients develop dilated cardiomyopathy. As the clinical picture of myocarditis is multifaceted, it is difficult to diagnose the disease. Therefore, the main goal of the present work was to test and further develop novel non-invasive methods for the detection of myocardial inflammation by employing both contrast enhanced MRI techniques as well as novel nuclear tracers that are suitable for in vivo PET/ SPECT imaging. As a part of this thesis, a pre-clinical animal model was successfully established by immunizing female Lewis rats with whole-porcine cardiac myosin (CM). Induction of Experimental Autoimmune Myocarditis (EAM) is considered successful when anti-myosin antibody titers are increased more than 100-fold over control animals and pericardial effusion develops. In addition, cardiac tissues from EAM-rats versus controls were analyzed for the expression of various pro-inflammatory and fibrosis markers. To further exploit non-invasive MRI techniques for the detection of myocarditis, our EAM-rats were injected either with (1) ultra-small Paramagnetic iron oxide particles (USPIO’s; Feraheme®), allowing for in vivo imaging , (2) micron sized paramagnetic iron oxide particles (MPIO) for ex vivo inflammatory cell-tracking by cMRI, or (3) with different radioactive nuclear tracers (67gallium citrate, 68gallium-labeled somatostatin analogue, and 68gallium-labeled cyclic RGD-peptide) which in the present work have been employed for autoradiographic imaging, but in principle are also suitable for in vivo nuclear imaging (PET/SPECT). In order to compare imaging results with histology, consecutive heart sections were stained with hematoxylin & eosin (HE, for cell infiltrates) and Masson Goldner trichrome (MGT, for fibrosis); in addition, immuno-stainings were performed with anti-CD68 (macrophages), anti-SSRT2A (somatostatin receptor type 2A), anti-CD61 (β3-integrins) and anti-CD31 (platelet endothelial cell adhesion molecule 1). Sera from immunized rats strongly reacted with cardiac myosin. In immunized rats, echocardiography and subsequent MRI revealed huge amounts of pericardial effusion (days 18-21). Analysis of the kinetics of myocardial infiltrates revealed maximal macrophage invasion between days 14 and 28. Disappearance of macrophages resulted in replacement-fibrosis in formerly cell-infiltrated myocardial areas. This finding was confirmed by the time-dependent differential expression of corresponding cytokines in the myocardium. Immunized animals reacted either with an early or a late pattern of post-inflammation fibrosis. Areas with massive cellular infiltrates were easily detectible in autoradiograms showing a high focal uptake of 67gallium-citrate and 68gallium labeled somatostatin analogues (68Ga DOTA-TATE). Myocardium with a loss of cardiomyocytes presented a high uptake of 68gallium labeled cyclic RGD-peptide (68Ga NOTA-RGD). MRI cell tracking experiments with Feraheme® as the contrast-agent were inconclusive; however, strikingly better results were obtained when MPIOs were used as a contrast-agent: histological findings correlated well with in vivo and ex vivo MPIO-enhanced MRI images. Imaging of myocardial inflammatory processes including the kinetics of macrophage invasion after microbial or ischemic damage is still a major challenge in, both animal models and in human patients. By applying a broad panel of biochemical, histological, molecular and imaging methods, we show here that different patterns of reactivity may occur upon induction of myocarditis using one and the same rat strain. In particular, immunized Lewis rats may react either with an early or a late pattern of macrophage invasion and subsequent post-inflammation fibrosis. Imaging results achieved in the acute inflammatory phase of the myocarditis with MPIOs, 67gallium citrate and 68gallium linked to somatostatin will stimulate further development of contrast agents and radioactive-nuclear tracers for the non-invasive detection of acute myocarditis and in the near future perhaps even in human patients. N2 - Eine fulminant verlaufende Myokarditis ist eine seltene aber potentiell lebensbedrohliche Erkrankung. Akute oder chronische Myokarditis gehen generell mit einer starken Aktivierung des Immunsystems der Betroffenen einher. Zum einen ist dies notwendig, um das Herz durch Bekämpfung der Eindringlinge (z.B. Bakterien, Viren oder andere mikrobielle Erreger) zu schützen und/oder Heilungs- und Reparaturprozesse im geschädigten Myokard einzuleiten. Zum anderen kann eine unkontrollierte Aktivierung des Immunsystems aber auch zur Entstehung von (nicht immer vorteilhaften) auto-reaktiven Immunzellen führen. Eine Myokarditis oder entzündliche Kardiomyopathie ist charakterisiert durch fokale oder diffuse Infiltrate, Nekrose und/oder Apoptose der Myozyten und einen fortschreitenden fibrotischen Ersatz des Herzmuskelgewebes. Beim Menschen entwickeln etwa 30% der Myokarditis-Patienten eine dilatative Kardiomyopathie. Da das klinische Bild der Myokarditis sehr vielfältig sein kann, ist die Diagnosestellung dieser Erkrankung schwierig. Deshalb war es das Kernziel dieser Arbeit, nicht-invasive Methoden zum Nachweis myokardialer Entzündungen zu testen, und dabei neue Bildgebungsverfahren unter Einsatz von neuen MRT-Kontrastmitteln sowie neuen nuklearen Tracern, die auch für PET/SPECT geeignet wären, zu entwickeln. Diese Verfahren wurden von uns zunächst an einem human-analogen Ratten-Modell evaluiert, mit dem Ziel später evtl. auch einmal beim Menschen eingesetzt werden zu können. Für unser präklinisches Tiermodell wurden weibliche Lewis-Ratten mit kardialem Myosin aus Schweinen immunisiert. Die erfolgreiche Induktion einer „Experimentellen Autoimmunen Myokarditis (EAM)“ wurde durch einen signifikanten Anstieg der Anti-Myosin Antikörpertiter in immunisierten Tieren und die Ausbildung eines Perikardergusses (Echokardiographie) bestätigt. Zusätzlich wurde aus apikalem kardialem Gewebe RNA isoliert und die Expression verschiedener pro-inflammatorischer und pro-fibrotischer molekularer Marker untersucht. Um die Bildgebung mittels kontrast-verstärktem cMRT zu optimieren, wurden den Tieren entweder kleine Eisenoxid-Nanopartikel (Ultra small paramagnetic iron oxide particles, USPIO; Feraheme®), oder sog. ,,Micronsized paramagnetic iron oxide particles (MPIO)‘‘ für das Tracking inflammatorischer Zellen injiziert. Im daraufolgenden Schritt wurden radioaktive nukleare Tracer (67Gallium-Citrat, 68Gallium-markierte Somatostatin-Analoga und 68Gallium-markierte zyklische RGD-Peptide) injiziert, um dann Autoradiogramme von Herzschnitten zu gewinnen. KW - Ratte KW - inflammation KW - Myokarditis KW - Kernspintomografie KW - myocarditis KW - MRI KW - cardiovascular KW - Entzündung Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-122469 ER - TY - JOUR A1 - Schick, Martin Alexander A1 - Baar, Wolfgang A1 - Bruno, Raphael Romano A1 - Wollborn, Jakob A1 - Held, Christopher A1 - Schneider, Reinhard A1 - Flemming, Sven A1 - Schlegel, Nicolas A1 - Roewer, Norbert A1 - Neuhaus, Winfried A1 - Wunder, Christian T1 - Balanced hydroxyethylstarch (HES 130/0.4) impairs kidney function in-vivo without inflammation JF - PLoS One N2 - Volume therapy is a standard procedure in daily perioperative care, and there is an ongoing discussion about the benefits of colloid resuscitation with hydroxyethylstarch (HES). In sepsis HES should be avoided due to a higher risk for acute kidney injury (AKI). Results of the usage of HES in patients without sepsis are controversial. Therefore we conducted an animal study to evaluate the impact of 6% HES 130/0.4 on kidney integrity with sepsis or under healthy conditions Sepsis was induced by standardized Colon Ascendens Stent Peritonitis (sCASP). sCASP-group as well as control group (C) remained untreated for 24 h. After 18 h sCASP+HES group (sCASP+VOL) and control+HES (C+VOL) received 50 ml/KG balanced 6% HES (VOL) 130/0.4 over 6h. After 24h kidney function was measured via Inulin- and PAH-Clearance in re-anesthetized rats, and serum urea, creatinine (crea), cystatin C and Neutrophil gelatinase-associated lipocalin (NGAL) as well as histopathology were analysed. In vitro human proximal tubule cells (PTC) were cultured +/- lipopolysaccharid (LPS) and with 0.1–4.0% VOL. Cell viability was measured with XTT-, cell toxicity with LDH-test. sCASP induced severe septic AKI demonstrated divergent results regarding renal function by clearance or creatinine measure focusing on VOL. Soleley HES (C+VOL) deteriorated renal function without sCASP. Histopathology revealed significantly derangements in all HES groups compared to control. In vitro LPS did not worsen the HES induced reduction of cell viability in PTC cells. For the first time, we demonstrated, that application of 50 ml/KG 6% HES 130/0.4 over 6 hours induced AKI without inflammation in vivo. Severity of sCASP induced septic AKI might be no longer susceptible to the way of volume expansion KW - colloids KW - kidneys KW - histopathology KW - blood KW - creatinine KW - sepsis KW - urine KW - inflammation Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-126068 VL - 10 IS - 9 ER - TY - JOUR A1 - Loeffler, Claudia A1 - Loeffler, Jürgen A1 - Kobsar, Anna A1 - Speer, Christian P. A1 - Eigenthaler, Martin T1 - Septic Vs Colonizing Group B Streptococci Differentially Regulate Inflammation and Apoptosis in Human Coronary Artery Endothelial Cells - a Pilot Study JF - Journal of Pediatrics and Neonatal Care N2 - In this pilot study, we exemplify differences between a septic and a colonizing GBS strain during their interaction with Endothelial Cells by evaluating cytokine levels, surface and apoptosis-related molecules. These preliminary results indicate that in vitro infection using an exemplary septic GBS strain results in diminished activation of the innate immune response. KW - streptococci KW - apoptosis KW - inflammation KW - endothelial cells KW - innate immunity KW - early onset sepsis Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-125596 VL - 2 IS - 2 ER - TY - JOUR A1 - Wagner, Martin A1 - Ashby, Damien R. A1 - Kurtz, Caroline A1 - Alam, Ahsan A1 - Busbridge, Mark A1 - Raff, Ulrike A1 - Zimmermann, Josef A1 - Heuschmann, Peter U. A1 - Wanner, Christoph A1 - Schramm, Lothar T1 - Hepcidin-25 in diabetic chronic kidney disease is predictive for mortality and progression to end stage renal disease JF - PLoS One N2 - Background Anemia is common and is associated with impaired clinical outcomes in diabetic chronic kidney disease (CKD). It may be explained by reduced erythropoietin (EPO) synthesis, but recent data suggest that EPO-resistance and diminished iron availability due to inflammation contribute significantly. In this cohort study, we evaluated the impact of hepcidin-25—the key hormone of iron-metabolism—on clinical outcomes in diabetic patients with CKD along with endogenous EPO levels. Methods 249 diabetic patients with CKD of any stage, excluding end-stage renal disease (ESRD), were enrolled (2003–2005), if they were not on EPO-stimulating agent and iron therapy. Hepcidin-25 levels were measured by radioimmunoassay. The association of hepcidin-25 at baseline with clinical variables was investigated using linear regression models. All-cause mortality and a composite endpoint of CKD progression (ESRD or doubling of serum creatinine) were analyzed by Cox proportional hazards models. Results Patients (age 67 yrs, 53% male, GFR 51 ml/min, hemoglobin 131 g/L, EPO 13.5 U/L, hepcidin-25 62.0 ng/ml) were followed for a median time of 4.2 yrs. Forty-nine patients died (19.7%) and forty (16.1%) patients reached the composite endpoint. Elevated hepcidin levels were independently associated with higher ferritin-levels, lower EPO-levels and impaired kidney function (all p<0.05). Hepcidin was related to mortality, along with its interaction with EPO, older age, greater proteinuria and elevated CRP (all p<0.05). Hepcidin was also predictive for progression of CKD, aside from baseline GFR, proteinuria, low albumin- and hemoglobin-levels and a history of CVD (all p<0.05). Conclusions We found hepcidin-25 to be associated with EPO and impaired kidney function in diabetic CKD. Elevated hepcidin-25 and EPO-levels were independent predictors of mortality, while hepcidin-25 was also predictive for progression of CKD. Both hepcidin-25 and EPO may represent important prognostic factors of clinical outcome and have the potential to further define “high risk” populations in CKD. KW - diabetes mellitus KW - inflammation KW - type 2 diabetes KW - hemoglobin KW - chronic kidney disease KW - anemia KW - ferritin KW - proteinuria Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-125514 VL - 10 IS - 4 ER - TY - JOUR A1 - Schrewe, L. A1 - Lill, C. M. A1 - Liu, T. A1 - Salmen, A. A1 - Gerdes, L. A. A1 - Guillot-Noel, L. A1 - Akkad, D. A. A1 - Blaschke, P. A1 - Graetz, C. A1 - Hoffjan, S. A1 - Kroner, A. A1 - Demir, S. A1 - Böhme, A. A1 - Rieckmann, P. A1 - El Ali, A. A1 - Hagemann, N. A1 - Hermann, D. M. A1 - Cournu-Rebeix, I. A1 - Zipp, F. A1 - Kümpfel, T. A1 - Buttmann, M. A1 - Zettl, U. K. A1 - Fontaine, B. A1 - Bertram, L. A1 - Gold, R. A1 - Chan, A. T1 - Investigation of sex-specific effects of apolipoprotein E on severity of EAE and MS JF - Journal of Neuroinflammation N2 - Background: Despite pleiotropic immunomodulatory effects of apolipoprotein E (apoE) in vitro, its effects on the clinical course of experimental autoimmune encephalomyelitis (EAE) and multiple sclerosis (MS) are still controversial. As sex hormones modify immunomodulatory apoE functions, they may explain contentious findings. This study aimed to investigate sex-specific effects of apoE on disease course of EAE and MS. Methods: MOG\(_{35-55}\) induced EAE in female and male apoE-deficient mice was assessed clinically and histopathologically. apoE expression was investigated by qPCR. The association of the MS severity score (MSSS) and APOE rs429358 and rs7412 was assessed across 3237 MS patients using linear regression analyses. Results: EAE disease course was slightly attenuated in male apoE-deficient (apoE\(^{-/-}\)) mice compared to wildtype mice (cumulative median score: apoE\(^{-/-}\) = 2 [IQR 0.0-4.5]; wildtype = 4 [IQR 1.0-5.0]; n = 10 each group, p = 0.0002). In contrast, EAE was more severe in female apoE\(^{-/-}\) mice compared to wildtype mice (cumulative median score: apoE\(^{-/-}\) = 3 [IQR 2.0-4.5]; wildtype = 3 [IQR 0.0-4.0]; n = 10, p = 0.003). In wildtype animals, apoE expression during the chronic EAE phase was increased in both females and males (in comparison to naive animals; p < 0.001). However, in MS, we did not observe a significant association between MSSS and rs429358 or rs7412, neither in the overall analyses nor upon stratification for sex. Conclusions: apoE exerts moderate sex-specific effects on EAE severity. However, the results in the apoE knock-out model are not comparable to effects of polymorphic variants in the human APOE gene, thus pinpointing the challenge of translating findings from the EAE model to the human disease. KW - immune KW - apoE KW - gender KW - inflammation KW - association studies in genetics KW - apoe KW - CNS disease KW - system KW - multiple sclerosis KW - MSSS KW - experimental autoimmune encephalomyelitis KW - disease severity KW - cognitive function KW - Alzheimer disease Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-136252 VL - 12 IS - 234 ER - TY - JOUR A1 - Schuhmann, Michael K. A1 - Bittner, Stefan A1 - Meuth, Sven G. A1 - Kleinschnitz, Christoph A1 - Fluri, Felix T1 - Fingolimod (FTY720-P) does not stabilize the blood-brain barrier under inflammatory conditions in an in vitro model JF - International Journal of Molecular Sciences N2 - Breakdown of the blood-brain barrier (BBB) is an early hallmark of multiple sclerosis (MS), a progressive inflammatory disease of the central nervous system. Cell adhesion in the BBB is modulated by sphingosine-1-phosphate (S1P), a signaling protein, via S1P receptors (S1P\(_1\)). Fingolimod phosphate (FTY720-P) a functional S1P\(_1\) antagonist has been shown to improve the relapse rate in relapsing-remitting MS by preventing the egress of lymphocytes from lymph nodes. However, its role in modulating BBB permeabilityin particular, on the tight junction proteins occludin, claudin 5 and ZO-1has not been well elucidated to date. In the present study, FTY720-P did not change the transendothelial electrical resistance in a rat brain microvascular endothelial cell (RBMEC) culture exposed to inflammatory conditions and thus did not decrease endothelial barrier permeability. In contrast, occludin was reduced in RBMEC culture after adding FTY720-P. Additionally, FTY720-P did not alter the amount of endothelial matrix metalloproteinase (MMP)-9 and MMP-2 in RBMEC cultures. Taken together, our observations support the assumption that S1P\(_1\) plays a dual role in vascular permeability, depending on its ligand. Thus, S1P\(_1\) provides a mechanistic basis for FTY720-P-associated disruption of endothelial barrierssuch as the blood-retinal barrierwhich might result in macular edema. KW - randomized controlled trial KW - Sphingosine 1-Phosphate KW - vascular permeability KW - rat brain microvascular endothelial cell culture KW - tight junctions KW - FTY720-P KW - blood-brain barrier KW - inflammation KW - novo renal transplantation KW - endothelial cells KW - experimental autoimmune encephalomyelitis KW - relapsing multiple sclerosis KW - Zonula Occludens-1 KW - matrix metalloproteinases Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-145047 VL - 16 ER - TY - JOUR A1 - Ebert, Regina A1 - Benisch, Peggy A1 - Krug, Melanie A1 - Zeck, Sabine A1 - Meißner-Weigl, Jutta A1 - Steinert, Andre A1 - Rauner, Martina A1 - Hofbauer, Lorenz A1 - Jakob, Franz T1 - Acute phase serum amyloid A induces proinflammatory cytokines and mineralization via toll-like receptor 4 in mesenchymal stem cells JF - Stem Cell Research N2 - The role of serum amyloid A (SAA) proteins, which are ligands for toll-like receptors, was analyzed in human bone marrow-derived mesenchymal stem cells (hMSCs) and their osteogenic offspring with a focus on senescence, differentiation andmineralization. In vitro aged hMSC developed a senescence-associated secretory phenotype (SASP), resulting in enhanced SAA1/2, TLR2/4 and proinflammatory cytokine (IL6, IL8, IL1\(\beta\), CXCL1, CXCL2) expression before entering replicative senescence. Recombinant human SAA1 (rhSAA1) induced SASP-related genes and proteins in MSC, which could be abolished by cotreatment with the TLR4-inhibitor CLI-095. The same pattern of SASP-resembling genes was stimulated upon induction of osteogenic differentiation, which is accompanied by autocrine SAA1/2 expression. In this context additional rhSAA1 enhanced the SASP-like phenotype, accelerated the proinflammatory phase of osteogenic differentiation and enhanced mineralization. Autocrine/paracrine and rhSAA1 via TLR4 stimulate a proinflammatory phenotype that is both part of the early phase of osteogenic differentiation and the development of senescence. This signaling cascade is tightly involved in bone formation and mineralization, but may also propagate pathological extraosseous calcification conditions such as calcifying inflammation and atherosclerosis. KW - human atherosclerotic lesions KW - senescence KW - expression KW - toll-like receptor KW - mineralization KW - osteogenic differentiation KW - serum amyloid A KW - inflammation KW - mesenchymal stem cells KW - WNT5A KW - model KW - lines KW - stromal cells KW - RT-PCR Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-148491 VL - 15 ER - TY - JOUR A1 - Macedo, Robson A1 - Javadi, Som Mehrbod A1 - Higuchi, Takahiro A1 - Ferreira de Carvalho, Marilia Daniela A1 - Lima Paiva Medeiros, Vanessa de Fátima A1 - Azevedo, Ítalo Medeiros A1 - Lima, Francisco Pignataro A1 - Medeiros, Aldo Cunha T1 - Heart and systemic effects of statin pretreatment in a rat model of abdominal sepsis. Assessment by Tc\(^{99m}\)-sestamibi biodistribition JF - Acta Cirúrgica Brasileira N2 - PURPOSE: To evaluate the heart and the Tc-99m-sestamibi biodistribution after statin pretreatment in a rat model of abdominal sepsis. METHODS: Twenty-four Wistar rats were randomly distributed into four groups (n=6 per group): 1) sepsis with simvastatin treatment, 2) sepsis with vehicle, 3) sham control with simvastatin and 4) sham control with vehicle. 24 hours after cecal ligation and puncture rats received 1.0MBq of Tc-99m-sestamibi i.v. 30min after, animals were euthanized for ex-vivo tissue counting and myocardium histological analysis. RESULTS: Myocardial histologic alterations were not detected 24 hours post-sepsis. There was significantly increased cardiac Tc-99m-sestamibi activity in the sepsis group with simvastatin treatment (1.9\(\pm\)0.3%ID/g, p<0.001) in comparison to the sepsis group+vehicle (1.0\(\pm\)0.2% ID/g), control sham group+ simvastatin (1.2\(\pm\)0.3% ID/g) and control sham group (1.3\(\pm\)0.2% ID/g). Significant Tc-99m-sestamibi activity in liver, kidney and lungs was also detected in the sepsis group treated with simvastatinin comparison to the other groups. CONCLUSIONS: Statin treatment altered the biodistribution of Tc-99m-sestamibi with increased cardiac and solid organ activity in rats with abdominal sepsis, while no impact on controls. Increased myocardial tracer activity may be a result of a possible protection effect due to increased tissue perfusion mediated by statins. KW - P-glycoprotein expression KW - mechanisms retention KW - Simvastatin KW - Technetium Tc 99m Sestamibi Rats KW - heart KW - inflammation KW - sepsis Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-151887 VL - 30 IS - 6 SP - 388 EP - 393 ER - TY - JOUR A1 - Albert-Weissenberger, Christiane A1 - Mencl, Stine A1 - Schuhmann, Michael K. A1 - Salur, Irmak A1 - Göb, Eva A1 - Langhauser, Friederike A1 - Hopp, Sarah A1 - Hennig, Nelli A1 - Meuth, Sven G. A1 - Nolte, Marc W. A1 - Sirén, Anna-Leena A1 - Kleinschnitz, Christoph T1 - C1-Inhibitor protects from focal brain trauma in a cortical cryolesion mice model by reducing thrombo-inflammation JF - Frontiers in Cellular Neuroscience N2 - Traumatic brain injury (TBI) induces a strong inflammatory response which includes blood-brain barrier damage, edema formation and infiltration of different immune cell subsets. More recently, microvascular thrombosis has been identified as another pathophysiological feature of TBI. The contact-kinin system represents an interface between inflammatory and thrombotic circuits and is activated in different neurological diseases. C1-Inhibitor counteracts activation of the contact-kinin system at multiple levels. We investigated the therapeutic potential of C1-Inhibitor in a model of TBI. Male and female C57BL/6 mice were subjected to cortical cryolesion and treated with C1-Inhibitor after 1 h. Lesion volumes were assessed between day 1 and day 5 and blood-brain barrier damage, thrombus formation as well as the local inflammatory response were determined post TBI. Treatment of male mice with 15.0 IU C1-Inhibitor, but not 7.5 IU, 1 h after cryolesion reduced lesion volumes by ~75% on day 1. This protective effect was preserved in female mice and at later stages of trauma. Mechanistically, C1-Inhibitor stabilized the blood-brain barrier and decreased the invasion of immune cells into the brain parenchyma. Moreover, C1-Inhibitor had strong antithrombotic effects. C1-Inhibitor represents a multifaceted anti-inflammatory and antithrombotic compound that prevents traumatic neurodegeneration in clinically meaningful settings. KW - thrombosis KW - traumatic brain injury KW - C1-inhibitor KW - blood-brain barrier KW - contact-kinin system KW - edema KW - inflammation Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-119263 SN - 1662-5102 VL - 8 ER - TY - JOUR A1 - Hütten, Mareike A1 - Dhanasingh, Anandhan A1 - Hessler, Roland A1 - Stöver, Timo A1 - Esser, Karl-Heinz A1 - Möller, Martin A1 - Lenarz, Thomas A1 - Jolly, Claude A1 - Groll, Jürgen A1 - Scheper, Verena T1 - In Vitro and In Vivo Evaluation of a Hydrogel Reservoir as a Continuous Drug Delivery System for Inner Ear Treatment JF - PLoS ONE N2 - Fibrous tissue growth and loss of residual hearing after cochlear implantation can be reduced by application of the glucocorticoid dexamethasone-21-phosphate-disodium-salt (DEX). To date, sustained delivery of this agent to the cochlea using a number of pharmaceutical technologies has not been entirely successful. In this study we examine a novel way of continuous local drug application into the inner ear using a refillable hydrogel functionalized silicone reservoir. A PEG-based hydrogel made of reactive NCO-sP(EO-stat-PO) prepolymers was evaluated as a drug conveying and delivery system in vitro and in vivo. Encapsulating the free form hydrogel into a silicone tube with a small opening for the drug diffusion resulted in delayed drug release but unaffected diffusion of DEX through the gel compared to the free form hydrogel. Additionally, controlled DEX release over several weeks could be demonstrated using the hydrogel filled reservoir. Using a guinea-pig cochlear trauma model the reservoir delivery of DEX significantly protected residual hearing and reduced fibrosis. As well as being used as a device in its own right or in combination with cochlear implants, the hydrogel-filled reservoir represents a new drug delivery system that feasibly could be replenished with therapeutic agents to provide sustained treatment of the inner ear. KW - gels KW - cochlea KW - silicones KW - deafness KW - inner ear KW - drug delivery KW - inflammation KW - connective tissue Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-119375 VL - 9 IS - 8 ER - TY - JOUR A1 - Cardani, Diego A1 - Sardi, Claudia A1 - La Ferla, Barbara A1 - D'Orazio, Guiseppe A1 - Sommariva, Michele A1 - Marcucci, Fabrizio A1 - Olivero, Daniela A1 - Tagliabue, Elda A1 - Koepsell, Hermann A1 - Nicotra, Francesco A1 - Balsari, Andrea A1 - Rumio, Christiano T1 - Sodium glucose cotransporter 1 ligand BLF501 as a novel tool for management of gastrointestinal mucositis JF - Molecular Cancer N2 - Background: Recent studies demonstrated that engagement of sodium glucose transporter 1 (SGLT-1) by orally administered D-glucose protects the intestinal mucosa from lipopolysaccharide (LPS)-induced injury. We tested whether SGLT-1 engagement might protect the intestinal mucosa from doxorubicin (DXR)- and 5-fluorouracil (5-FU)-induced injury in animal models mimicking acute or chronic mucositis. Methods: Mice were treated intraperitoneally with DXR, alone or in combination with 5-FU, and orally with BLF501, a glucose-derived synthetic compound with high affinity for SGLT-1. Intestinal mucosal epithelium integrity was assessed by histological analysis, cellular proliferation assays, real-time PCR gene expression assays and Western blot assays. Student's t-test (paired two-tailed) and X-2 analyses were used for comparisons between groups. Differences were considered significant at p < 0.05. Results: BLF501 administration in mice treated with DXR and/or 5-FU decreased the injuries to the mucosa in terms of epithelial integrity and cellular proliferative ability. Co-treatment with BLF501 led to a normal expression and distribution of both zonula occludens-1 (ZO-1) and beta-catenin, which were underexpressed after treatment with either chemotherapeutic agent alone. BLF501 administration also restored normal expression of caspase-3 and ezrin/radixin/moesin (ERM), which were overexpressed after treatment with DXR and 5-FU. In SGLT1-/- mice, BLF501 had no detectable effects. BLF501 administration in wild-type mice with growing A431 tumors did not modify antitumor activity of DXR. Conclusions: BLF501-induced protection of the intestinal mucosa is a promising novel therapeutic approach to reducing the severity of chemotherapy-induced mucositis. KW - apoptosis KW - prevention KW - doxorubicin KW - cancer KW - gastrointestinal mucositis KW - SGLT-1 KW - synthetic D-glucose analogy KW - chemotherapy KW - inflammation KW - clinical practice guidelines KW - intestinal mucositis KW - epithelial cells KW - oral mucositis KW - gene-expression Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-117352 SN - 1476-4598 VL - 13 IS - 23 ER - TY - JOUR A1 - Rittner, Heike L. A1 - Wang, Ying A1 - Gehringer, Rebekka A1 - Mousa, Shaaban A. A1 - Hackel, Dagmar A1 - Brack, Alexander T1 - CXCL10 Controls Inflammatory Pain via Opioid Peptide- Containing Macrophages in Electroacupuncture N2 - Acupuncture is widely used for pain treatment in patients with osteoarthritis or low back pain, but molecular mechanisms remain largely enigmatic. In the early phase of inflammation neutrophilic chemokines direct opioid-containing neutrophils in the inflamed tissue and stimulate opioid peptide release and antinociception. In this study the molecular pathway and neuroimmune connections in complete Freund's adjuvant (CFA)-induced hind paw inflammation and electroacupuncture for peripheral pain control were analyzed. Free moving Wistar rats with hind paw inflammation were treated twice with electroacupuncture at GB30 (Huan Tiao - gall bladder meridian) (day 0 and 1) and analyzed for mechanical and thermal nociceptive thresholds. The cytokine profiles as well as the expression of opioid peptides were quantified in the inflamed paw. Electroacupuncture elicited long-term antinociception blocked by local injection of anti-opioid peptide antibodies (beta-endorphin, met-enkephalin, dynorphin A). The treatment altered the cytokine profile towards an anti-inflammatory pattern but augmented interferon (IFN)-gamma and the chemokine CXCL10 (IP-10: interferon gamma-inducible protein) protein and mRNA expression with concomitant increased numbers of opioid peptide-containing CXCR3+ macrophages. In rats with CFA hind paw inflammation without acupuncture repeated injection of CXCL10 triggered opioid-mediated antinociception and increase opioid-containing macrophages. Conversely, neutralization of CXCL10 time-dependently decreased electroacupuncture-induced antinociception and the number of infiltrating opioid peptide-expressing CXCR3+ macrophages. In summary, we describe a novel function of the chemokine CXCL10 - as a regulator for an increase of opioid-containing macrophages and antinociceptive mediator in inflammatory pain and as a key chemokine regulated by electroacupuncture. KW - opioids KW - inflammation KW - macrophages KW - cytokines KW - chemokines KW - enzyme-linkes immunoassays KW - acupuncture KW - analysis of variance Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-112979 ER - TY - JOUR A1 - Hedrich, Christian M. A1 - Hofmann, Sigrun R. A1 - Pablik, Jessica A1 - Morbach, Henner A1 - Girschick, Hermann J. T1 - Autoinflammatory bone disorders with special focus on chronic recurrent multifocal osteomyelitis (CRMO) JF - Pediatric Rheumatology N2 - Sterile bone inflammation is the hallmark of autoinflammatory bone disorders, including chronic nonbacterial osteomyelitis (CNO) with its most severe form chronic recurrent multifocal osteomyelitis (CRMO). Autoinflammatory osteopathies are the result of a dysregulated innate immune system, resulting in immune cell infiltration of the bone and subsequent osteoclast differentiation and activation. Interestingly, autoinflammatory bone disorders are associated with inflammation of the skin and/or the intestine. In several monogenic autoinflammatory bone disorders mutations in disease-causing genes have been reported. However, regardless of recent developments, the molecular pathogenesis of CNO/CRMO remains unclear. Here, we discuss the clinical presentation and molecular pathophysiology of human autoinflammatory osteopathies and animal models with special focus on CNO/CRMO. Treatment options in monogenic autoinflammatory bone disorders and CRMO will be illustrated. KW - bisphosphonate treatment KW - IL-10 expression KW - TNF-α KW - IL-10 KW - inflammation KW - bone KW - CRMO KW - CNO KW - DIRA KW - PAPA KW - Majeed-Syndrome KW - disease KW - deficiency KW - pediatric patients KW - treatment KW - TLR4 KW - PAPA syndrome KW - hypertrophic osteodystrophy KW - chronic nonbacterial osteomyelitis KW - congenital dyserythropoietic anemia Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-125694 SN - 1546-0096 VL - 11 IS - 47 ER - TY - JOUR A1 - Blömer, Nadja A1 - Pachel, Christina A1 - Hofmann, Urlich A1 - Nordbeck, Peter A1 - Bauer, Wolfgang A1 - Mathes, Denise A1 - Frey, Anna A1 - Bayer, Barbara A1 - Vogel, Benjamin A1 - Ertl, Georg T1 - 5-Lipoxygenase facilitates healing after myocardial infarction JF - Basic Research in Cardiology N2 - Early healing after myocardial infarction (MI) is characterized by a strong inflammatory reaction. Most leukotrienes are pro-inflammatory and are therefore potential mediators of healing and remodeling after myocardial ischemia. The enzyme 5-lipoxygenase (5-LOX) has a key role in the transformation of arachidonic acid in leukotrienes. Thus, we tested the effect of 5-LOX on healing after MI. After chronic coronary artery ligation, early mortality was significantly increased in 5-LOX\(^{−/−}\) when compared to matching wildtype (WT) mice due to left ventricular rupture. This effect could be reproduced in mice treated with the 5-LOX inhibitor Zileuton. A perfusion mismatch due to the vasoactive potential of leukotrienes is not responsible for left ventricular rupture since local blood flow assessed by magnetic resonance perfusion measurements was not different. However, after MI, there was an accentuation of the inflammatory reaction with an increase of pro-inflammatory macrophages. Yet, mortality was not changed in chimeric mice (WT vs. 5-LOX\(^{−/−}\) bone marrow in 5-LOX\(^{−/−}\) animals), indicating that an altered function of 5-LOX\(^{−/−}\) inflammatory cells is not responsible for the phenotype. Collagen production and accumulation of fibroblasts were significantly reduced in 5-LOX\(^{−/−}\) mice in vivo after MI. This might be due to an impaired migration of 5-LOX\(^{−/−}\) fibroblasts, as shown in vitro to serum. In conclusion, a lack or inhibition of 5-LOX increases mortality after MI because of healing defects. This is not mediated by a change in local blood flow, but through an altered inflammation and/or fibroblast function. KW - lipoxygenase KW - myocardial infarction KW - extracellular matrix remodeling KW - inflammation Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-132602 VL - 108 IS - 4 ER - TY - JOUR A1 - Hedrich, Christian M. A1 - Hofmann, Sigrun R. A1 - Pablik, Jessica A1 - Morbach, Henner A1 - Girschick, Hermann J. T1 - Autoinflammatory bone disorders with special focus on chronic recurrent multifocal osteomyelitis (CRMO) JF - Pediatric Rheumatology N2 - Sterile bone inflammation is the hallmark of autoinflammatory bone disorders, including chronic nonbacterial osteomyelitis (CNO) with its most severe form chronic recurrent multifocal osteomyelitis (CRMO). Autoinflammatory osteopathies are the result of a dysregulated innate immune system, resulting in immune cell infiltration of the bone and subsequent osteoclast differentiation and activation. Interestingly, autoinflammatory bone disorders are associated with inflammation of the skin and/or the intestine. In several monogenic autoinflammatory bone disorders mutations in disease-causing genes have been reported. However, regardless of recent developments, the molecular pathogenesis of CNO/CRMO remains unclear. Here, we discuss the clinical presentation and molecular pathophysiology of human autoinflammatory osteopathies and animal models with special focus on CNO/CRMO. Treatment options in monogenic autoinflammatory bone disorders and CRMO will be illustrated. KW - TNF-α KW - PAPA KW - DIRA KW - Majeed KW - CNO KW - CRMO KW - bone KW - inflammation KW - IL-10 KW - treatment KW - TLR4 Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-132456 VL - 11 IS - 47 ER - TY - JOUR A1 - Weibel, Stephanie A1 - Basse-Luesebrink, Thomas Christian A1 - Hess, Michael A1 - Hofmann, Elisabeth A1 - Seubert, Carolin A1 - Langbein-Laugwitz, Johanna A1 - Gentschev, Ivaylo A1 - Sturm, Volker Jörg Friedrich A1 - Ye, Yuxiang A1 - Kampf, Thomas A1 - Jakob, Peter Michael A1 - Szalay, Aladar A. T1 - Imaging of Intratumoral Inflammation during Oncolytic Virotherapy of Tumors by \(^{19}\)F-Magnetic Resonance Imaging (MRI) JF - PLoS ONE N2 - Background Oncolytic virotherapy of tumors is an up-coming, promising therapeutic modality of cancer therapy. Unfortunately, non-invasive techniques to evaluate the inflammatory host response to treatment are rare. Here, we evaluate \(^{19}\)F magnetic resonance imaging (MRI) which enables the non-invasive visualization of inflammatory processes in pathological conditions by the use of perfluorocarbon nanoemulsions (PFC) for monitoring of oncolytic virotherapy. Methodology/Principal Findings The Vaccinia virus strain GLV-1h68 was used as an oncolytic agent for the treatment of different tumor models. Systemic application of PFC emulsions followed by \(^1H\)/\(^{19}\)F MRI of mock-infected and GLV-1h68-infected tumor-bearing mice revealed a significant accumulation of the \(^{19}\)F signal in the tumor rim of virus-treated mice. Histological examination of tumors confirmed a similar spatial distribution of the \(^{19}\)F signal hot spots and \(CD68^+\)-macrophages. Thereby, the \(CD68^+\)-macrophages encapsulate the GFP-positive viral infection foci. In multiple tumor models, we specifically visualized early inflammatory cell recruitment in Vaccinia virus colonized tumors. Furthermore, we documented that the \(^{19}\)F signal correlated with the extent of viral spreading within tumors. Conclusions/Significance These results suggest \(^{19}\)F MRI as a non-invasive methodology to document the tumor-associated host immune response as well as the extent of intratumoral viral replication. Thus, \(^{19}\)F MRI represents a new platform to non-invasively investigate the role of the host immune response for therapeutic outcome of oncolytic virotherapy and individual patient response. KW - inflammation KW - fluorescence microscopy KW - oncolytic viruses KW - fluorescence imaging KW - macrophages KW - magnetic resonance imaging KW - histology KW - in vivo imaging Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-130311 VL - 8 IS - 3 ER - TY - JOUR A1 - Wolf, Annette A1 - Akrap, Nina A1 - Marg, Berenice A1 - Galliardt, Helena A1 - Heiligentag, Martyna A1 - Humpert, Fabian A1 - Sauer, Markus A1 - Kaltschmidt, Barbara A1 - Kaltschmidt, Christian A1 - Seidel, Thorsten T1 - Elements of Transcriptional Machinery Are Compatible among Plants and Mammals JF - PLoS ONE N2 - In the present work, the objective has been to analyse the compatibility of plant and human transcriptional machinery. The experiments revealed that nuclear import and export are conserved among plants and mammals. Further it has been shown that transactivation of a human promoter occurs by human transcription factor NF-\(\kappa\) B in plant cells, demonstrating that the transcriptional machinery is highly conserved in both kingdoms. Functionality was also seen for regulatory elements of NF-\(\kappa\) B such as its inhibitor I\(\kappa\)B isoform \(\alpha\) that negatively regulated the transactivation activity of the p50/RelA heterodimer by interaction with NF-\(\kappa\)B in plant cells. Nuclear export of RelA could be demonstrated by FRAP-measurements so that RelA shows nucleo-cytoplasmic shuttling as reported for RelA in mammalian cells. The data reveals the high level of compatibility of human transcriptional elements with the plant transcriptional machinery. Thus, Arabidopsis thaliana mesophyll protoplasts might provide a new heterologous expression system for the investigation of the human NF-\(\kappa\)B signaling pathways. The system successfully enabled the controlled manipulation of NF-\(\kappa\)B activity. We suggest the plant protoplast system as a tool for reconstitution and analyses of mammalian pathways and for direct observation of responses to e. g. pharmaceuticals. The major advantage of the system is the absence of interference with endogenous factors that affect and crosstalk with the pathway. KW - complexes KW - in vivo KW - DNA-binding KW - nuclear proe KW - gene expression KW - NF-KAPPA-B KW - RNA-binding protein KW - alpha KW - inflammation KW - homodimers Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-131203 VL - 8 IS - 1 ER - TY - JOUR A1 - Pachel, Christina A1 - Mathes, Denise A1 - Bayer, Barbara A1 - Dienesch, Charlotte A1 - Wangorsch, Gaby A1 - Heitzmann, Wolfram A1 - Lang, Isabell A1 - Ardehali, Hossein A1 - Ertl, Georg A1 - Dandekar, Thomas A1 - Wajant, Harald A1 - Frantz, Stefan T1 - Exogenous Administration of a Recombinant Variant of TWEAK Impairs Healing after Myocardial Infarction by Aggravation of Inflammation JF - PLoS ONE N2 - Background: Tumor necrosis factor-like weak inducer of apoptosis (TWEAK) and its receptor fibroblast growth factorinducible 14 (Fn14) are upregulated after myocardial infarction (MI) in both humans and mice. They modulate inflammation and the extracellular matrix, and could therefore be important for healing and remodeling after MI. However, the function of TWEAK after MI remains poorly defined. Methods and results: Following ligation of the left coronary artery, mice were injected twice per week with a recombinant human serum albumin conjugated variant of TWEAK (HSA-Flag-TWEAK), mimicking the activity of soluble TWEAK. Treatment with HSA-Flag-TWEAK resulted in significantly increased mortality in comparison to the placebo group due to myocardial rupture. Infarct size, extracellular matrix remodeling, and apoptosis rates were not different after MI. However, HSA-Flag-TWEAK treatment increased infiltration of proinflammatory cells into the myocardium. Accordingly, depletion of neutrophils prevented cardiac ruptures without modulating all-cause mortality. Conclusion: Treatment of mice with HSA-Flag-TWEAK induces myocardial healing defects after experimental MI. This is mediated by an exaggerated neutrophil infiltration into the myocardium. KW - apoptosis KW - myocardial infarction KW - neutrophils KW - cytokines KW - inflammation KW - myocardium KW - heart KW - extracellular matrix Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-129889 VL - 8 IS - 11 ER - TY - JOUR A1 - Rittner, Heike Lydia A1 - Hackel, Dagmar A1 - Pflücke, Diana A1 - Neumann, Annick A1 - Viebahn, Johannes A1 - Mousa, Shaaban A1 - Wischmeyer, Erhard A1 - Roewer, Norbert A1 - Brack, Alexander T1 - The Connection of Monocytes and Reactive Oxygen Species in Pain JF - PLoS ONE N2 - The interplay of specific leukocyte subpopulations, resident cells and proalgesic mediators results in pain in inflammation. Proalgesic mediators like reactive oxygen species (ROS) and downstream products elicit pain by stimulation of transient receptor potential (TRP) channels. The contribution of leukocyte subpopulations however is less clear. Local injection of neutrophilic chemokines elicits neutrophil recruitment but no hyperalgesia in rats. In meta-analyses the monocytic chemoattractant, CCL2 (monocyte chemoattractant protein-1; MCP-1), was identified as an important factor in the pathophysiology of human and animal pain. In this study, intraplantar injection of CCL2 elicited thermal and mechanical pain in Wistar but not in Dark Agouti (DA) rats, which lack p47phox, a part of the NADPH oxidase complex. Inflammatory hyperalgesia after complete Freund's adjuvant (CFA) as well as capsaicin-induced hyperalgesia and capsaicin-induced current flow in dorsal root ganglion neurons in DA were comparable to Wistar rats. Macrophages from DA expressed lower levels of CCR2 and thereby migrated less towards CCL2 and formed limited amounts of ROS in vitro and 4-hydroxynonenal (4-HNE) in the tissue in response to CCL2 compared to Wistar rats. Local adoptive transfer of peritoneal macrophages from Wistar but not from DA rats reconstituted CCL2-triggered hyperalgesia in leukocyte-depleted DA and Wistar rats. A pharmacological stimulator of ROS production (phytol) restored CCL2-induced hyperalgesia in vivo in DA rats. In Wistar rats, CCL2-induced hyperalgesia was completely blocked by superoxide dismutase (SOD), catalase or tempol. Likewise, inhibition of NADPH oxidase by apocynin reduced CCL2-elicited hyperalgesia but not CFA-induced inflammatory hyperalgesia. In summary, we provide a link between CCL2, CCR2 expression on macrophages, NADPH oxidase, ROS and the development CCL2-triggered hyperalgesia, which is different from CFA-induced hyperalgesia. The study further supports the impact of CCL2 and ROS as potential targets in pain therapy. KW - analysis of variance KW - chemokines KW - hyperalgesia KW - inflammation KW - macrophages KW - monocytes KW - white blood cells KW - wistar rats Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-96669 ER - TY - THES A1 - Partheil, Anna T1 - Monozyten und Prostaglandine in der Schmerzentstehung T1 - Monocytes and prostaglandins in the development of inflammatory pain N2 - Schmerz ist eine klassische Komponente von Entzündungsreaktionen. Im Rahmen des Entzündungsgeschehens werden Zytokine und Chemokine freigesetzt, die Leukozyten zum Entzündungsort rekrutieren. Über die Freisetzung weiterer proalgetischer Mediatoren tragen diese zur Aktivierung und Sensitivierung von Nozizeptoren und damit zur Schmerzentstehung bei. Das Monozyten-rekrutierende Chemokin CCL2 verursachte in Verhaltensexperimenten eine Hyperalgesie bei Ratten. Die Hyperalgesie war durch den Cox-2 Inhibitor Parecoxib vollständig reversibel. Daher wurde in dieser Arbeit die Rolle von Monozyten und Prostaglandinen in der Entstehung dieser Hyperalgesie untersucht. Dazu wurde in vitro die Cox-2 Expression und die Prostaglandin-Bildung in humanen Monozyten und Peritonealmakrophagen der Ratte nach CCL2 Stimulation bestimmt. Zudem wurde in vivo die Cox-2 Expression im Rückenmark und in der Rattenpfote nach CCL2 Injektion in die Pfote untersucht. N2 - One of the main components of inflammation is pain. In inflammation cytokines and chemokines are secreted and recruit leukocytes to the site of inflammation. Leukocytes release proalgesic mediators that activate and sensitize nociceptors and cause pain. Behavioral tests showed that the chemokine CCL2 (monocyte recruiting protein 2) causes hyperalgesia in rats. This hyperalgesia was blocked by parecoxib, a Cox-2 inhibitor. To further investigate the role of monocytes and prostaglandins in the development of hyperalgesia, Cox-2 expression and prostaglandin production were determined in vitro after stimulation of human monocytes and rat peritoneal macrophages with CCL2. In vivo, Cox-2 expression in spinal cord and paw tissue of rats was examined after injection of CCL2 into the paw. KW - Schmerz KW - CCL2 KW - pain KW - Entzündung KW - Monozyten KW - Prostaglandine KW - inflammation KW - monocytes KW - prostaglandins Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-85091 ER - TY - JOUR A1 - Schupp, Nicole A1 - Ali, Badreldin H. A1 - Beegam, Sumyia A1 - Al-Husseni, Isehaq A1 - Al-Shukaili, Ahmed A1 - Nemmar, Abderrahim A1 - Schierling, Simone A1 - Queisser, Nina T1 - Effect of gum arabic on oxidative stress and inflammation in adenine-induced chronic renal failure in rats JF - PLoS One N2 - Inflammation and oxidative stress are known to be involved in the pathogenesis of chronic kidney disease in humans, and in chronic renal failure (CRF) in rats. The aim of this work was to study the role of inflammation and oxidative stress in adenine-induced CRF and the effect thereon of the purported nephroprotective agent gum arabic (GA). Rats were divided into four groups and treated for 4 weeks as follows: control, adenine in feed (0.75%, w/w), GA in drinking water (15%, w/v) and adenine+GA, as before. Urine, blood and kidneys were collected from the rats at the end of the treatment for analysis of conventional renal function tests (plasma creatinine and urea concentration). In addition, the concentrations of the pro-inflammatory cytokine TNF-a and the oxidative stress markers glutathione and superoxide dismutase, renal apoptosis, superoxide formation and DNA double strand break frequency, detected by immunohistochemistry for c-H2AX, were measured. Adenine significantly increased the concentrations of urea and creatinine in plasma, significantly decreased the creatinine clearance and induced significant increases in the concentration of the measured inflammatory mediators. Further, it caused oxidative stress and DNA damage. Treatment with GA significantly ameliorated these actions. The mechanism of the reported salutary effect of GA in adenine-induced CRF is associated with mitigation of the adenine-induced inflammation and generation of free radicals. KW - adenine KW - blood plasma KW - creatinine KW - inflammation KW - inflammatory diseases KW - Kidneys KW - Oxidative stress KW - Water resources Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-95787 ER - TY - JOUR A1 - Baune, Bernhard T. A1 - Konrad, Carsten A1 - Grotegerd, Dominik A1 - Suslow, Thomas A1 - Birosova, Eva A1 - Ohrmann, Patricia A1 - Bauer, Jochen A1 - Arolt, Volker A1 - Heindel, Walter A1 - Domschke, Katharina A1 - Schöning, Sonja A1 - Rauch, Astrid V. A1 - Uhlmann, Christina A1 - Kugel, Harald A1 - Dannlowski, Udo T1 - Interleukin-6 gene (IL-6): a possible role in brain morphology in the healthy adult brain JF - Journal of Neuroinflammation N2 - Background: Cytokines such as interleukin 6 (IL-6) have been implicated in dual functions in neuropsychiatric disorders. Little is known about the genetic predisposition to neurodegenerative and neuroproliferative properties of cytokine genes. In this study the potential dual role of several IL-6 polymorphisms in brain morphology is investigated. Methodology: In a large sample of healthy individuals (N = 303), associations between genetic variants of IL-6 (rs1800795; rs1800796, rs2069833, rs2069840) and brain volume (gray matter volume) were analyzed using voxel-based morphometry (VBM). Selection of single nucleotide polymorphisms (SNPs) followed a tagging SNP approach (e. g., Stampa algorigthm), yielding a capture 97.08% of the variation in the IL-6 gene using four tagging SNPs. Principal findings/results In a whole-brain analysis, the polymorphism rs1800795 (-174 C/G) showed a strong main effect of genotype (43 CC vs. 150 CG vs. 100 GG; x = 24, y = -10, z = -15; F(2,286) = 8.54, p(uncorrected) = 0.0002; p(AlphaSim-corrected) = 0.002; cluster size k = 577) within the right hippocampus head. Homozygous carriers of the G-allele had significantly larger hippocampus gray matter volumes compared to heterozygous subjects. None of the other investigated SNPs showed a significant association with grey matter volume in whole-brain analyses. Conclusions/significance: These findings suggest a possible neuroprotective role of the G-allele of the SNP rs1800795 on hippocampal volumes. Studies on the role of this SNP in psychiatric populations and especially in those with an affected hippocampus (e.g., by maltreatment, stress) are warranted. KW - aging brain KW - hippocampal neurogenesis KW - cholinergic neurons KW - neurothrophic factor KW - Alzheimers disease KW - neurite outgrowth KW - inflammatory cytokines KW - major depression KW - nervour system KW - dentate gyrus KW - genetics KW - inflammation KW - interleukin 6 KW - neuroprotection KW - voxel-based morphometry Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-130804 VL - 9 IS - 125 ER - TY - JOUR A1 - Willems, Coen H. M. P. A1 - Urlichs, Florian A1 - Seidenspinner, Silvia A1 - Kunzmann, Steffen A1 - Speer, Christian P. A1 - Kramer, Boris W. T1 - Poractant alfa (Curosurf (R)) increases phagocytosis of apoptotic neutrophils by alveolar macrophages in vivo JF - Respiratory Research N2 - Background: Clearance of apoptotic neutrophils in the lung is an essential process to limit inflammation, since they could become a pro-inflammatory stimulus themselves. The clearance is partially mediated by alveolar macrophages, which phagocytose these apoptotic cells. The phagocytosis of apoptotic immune cells by monocytes in vitro has been shown to be augmented by several constituents of pulmonary surfactant, e. g. phospholipids and hydrophobic surfactant proteins. In this study, we assessed the influence of exogenous poractant alfa (Curosurf (R)) instillation on the in vivo phagocytosis of apoptotic neutrophils by alveolar macrophages. Methods: Poractant alfa (200 mg/kg) was instilled intratracheally in the lungs of three months old adult male C57/Black 6 mice, followed by apoptotic neutrophil instillation. Bronchoalveloar lavage was performed and alveolar macrophages and neutrophils were counted. Phagocytosis of apoptotic neutrophils was quantified by determining the number of apoptotic neutrophils per alveolar macrophages. Results: Exogenous surfactant increased the number of alveolar macrophages engulfing apoptotic neutrophils 2.6 fold. The phagocytosis of apoptotic neutrophils was increased in the presence of exogenous surfactant by a 4.7 fold increase in phagocytosed apoptotic neutrophils per alveolar macrophage. Conclusions: We conclude that the anti-inflammatory properties of surfactant therapy may be mediated in part by increased numbers of alveolar macrophages and increased phagocytosis of apoptotic neutrophils by alveolar macrophages. KW - preterm KW - surfactant protein-A KW - respiratory-distress-syndrome KW - synthetic surfactant KW - human monocytes KW - SIRP-alpha KW - lung KW - cells KW - inflammation KW - resolution KW - anti inflammation KW - drug therapy KW - surfactant Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-130721 VL - 13 IS - 17 ER - TY - THES A1 - Rodenberg, Hella Katharina T1 - Inflammation und Mangelernährung bei Dialysepatienten mit Diabetes Mellitus Typ 2 T1 - Inflammation and malnutrition in patients with type 2 diabtes mellitus on hemodialysis N2 - In der vorliegenden Arbeit wird der Einfluss von hs-CRP und Albumin auf die kardiovaskuläre Ereignisrate und das Überleben von Patienten mit Diabetes Mellitus Typ 2 an der Hämodialyse untersucht. Grundlagen für die hier dargestellte Auswertung sind die in der 4D Studie erhobenen Daten. Die 4D Studie hat prospektiv, randomisiert, doppelblind und placebo-kontrolliert untersucht, ob die Behandlung mit Atorvastatin bei Patienten mit Diabetes Mellitus Typ 2 an der Hämodialyse den primären Endpunkt bestehend aus Herzinfarkt, kardialem Tod und Schlaganfall zu senken vermag. Die Daten zum hs-CRP und Albumin wurden bei Studienbeginn und nach sechs Monaten erhoben. In einer post-hoc Analyse mit Hilfe eines multivariaten Cox Regressionsmodels konnte bestätigt werden, dass ein erhöhter Spiegel an hs-CRP und ein verminderter Spiegel an Albumin im Zusammenhang mit einer vermehrten kardiovaskulären Ereignisrate und Mortalität stehen. Eine Behandlung mit Atorvastatin führte zwar nicht zu einer Risikosenkung für den primären Endpunkt oder die Mortalität, hatte aber einen stabilisierenden Effekt auf des hs-CRP Spiegel. N2 - In this dissertation the influence of high sensitive (hs)-CRP and Albumin on cardiovascular (cv) events and mortality in patients with type 2 diabetes mellitus on hemodialysis is determined. Based on the data of the „4D“ (Die Deutsche Dialyse Diabetes) Study a prospective, double-blind, placebo controlled study a post-hoc analysis was accomplished. The data of hs-CRP and Albumin were aroused at baseline and six month later. Via multivariate cox-regressions analysis it could be considered that elevated hs-CRP and reduced Albumin are associated with an increased risk for cv events and mortality. A treatment with Atorvastatin indeed didn’t neither reduce the risk for cv events nor the risk for mortality but had a stabilizing effect on hs-CRP level. KW - Hämodialyse KW - Mangelernährung KW - Proteinmangel KW - Inflammation KW - hs-CRP KW - Albumin KW - inflammation KW - malnutrition KW - hemodialysis KW - hs-CRP KW - albumin Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-71145 ER - TY - THES A1 - Rauh, Katharina T1 - Erythropoietin und Inflammation bei diabetischer Nephropathie T1 - Erythropoietin and Inflammation in Diabetic Chronic Kidney Disease N2 - Bei Patienten mit Diabetes mellitus Typ 2 und chronischer Niereninsuffizienz ist Anämie häufig. Zum Teil ist sie durch ungenügende EPO-Produktion bedingt. Zusätzlich wird die Hämoglobinsynthese, wie bei der Anämie chronischer Krankheiten (anemia of chronic disease, ACD) beschrieben, durch entzündliche Vorgänge unterdrückt. Der Stellenwert endogenen Erythropoietins bei Patienten mit diabetischer Nephropathie und ACD bleibt noch unsicher sowie auch der Zusammenhang zwischen EPO und der Nierenfunktion. Diese Querschnittsanalyse schloss 224 Patienten mit Typ 2-Diabetes in allen Stadien chronischer Niereninsuffizienz (CNI-Stadium 1-5) ein. Das mediane Alter betrug 67 Jahre, 54 % waren männlich und die mediane GFR lag bei 49 ml/min. Gemäß den Definitionen der K/DOQI-Richtlinien waren 41 % der Patienten anämisch. Von der Studie ausgeschlossen wurden wegen der Anämie behandelte Patienten und solche mit Eisenmangel. Prädiktoren der log-transformierten EPO-Spiegel wurden unter Benutzung multivariater linearer Regressionsmodelle ausgewertet. Die univariate und inverse Beziehung zwischen GFR und EPO-Spiegeln (p = 0,009) wurde in der multivariaten Analyse nicht-signifikant. Erhöhtes CRP (p < 0,001), niedriges Ferritin (p = 0,002), kardiovaskuläre Ereignisse in der Vorgeschichte (p = 0,02) und Hypertension (p = 0,04) waren nach Adjustierung für Alter, Geschlecht, Hb, GFR und andere klinische Faktoren unabhängig mit erhöhten EPO-Spiegeln assoziiert. In der untersuchten Population fand sich kein Zusammenhang zwischen EPO-Spiegeln und Hämoglobin. Bei diabetischen Patienten mit chronischer Niereninsuffizienz werden die EPO-Spiegel trotz gleichzeitig niedriger Hämoglobinspiegel vor allem durch Entzündungsparameter und den Eisenstatus vorhergesagt und sind dabei unabhängig von der Nierenfunktion. Deshalb könnte die Anämie bei Patienten mit diabetischer Nephropathie hauptsächlich durch Inflammation entstehen, die zu einem relativen Eisenmangel und einer Resistenz des Knochenmarks gegenüber endogenem EPO führt. N2 - BACKGROUND: Anemia is prevalent in patients with type 2 diabetes mellitus (DM) and chronic kidney disease (CKD) and is caused in part by insufficient erythropoietin (EPO) production. In addition, inflammatory processes also suppress hemoglobin synthesis as described in anemia of chronic diseases (ACD). The role of endogenous EPO in diabetic CKD and in ACD and its relationship to kidney function remain uncertain. METHODS: This cross-sectional analysis included 224 diabetic patients with CKD stages 1 to 5 (median age 67 yrs, 54% male, median GFR 49 ml/min). Anemia was defined as per K/DOQI guidelines and was prevalent in 41% of the patients. Patients treated for anemia and iron deficient patients were excluded. Predictors of log-transformed EPO-levels were evaluated using multivariate linear regression modeling. RESULTS: The univariate and inverse relationship between GFR and EPO-level (p = 0.009) became non-significant in multivariate analyses. Elevated C-reactive protein (p < 0.001), low ferritin (p = 0.002) , history of CVD (p = 0.02) and hypertension (p = 0.04) were independently associated with elevated EPO-level, after adjusting for age, gender, hemoglobin, GFR, and other clinical factors. Hemoglobin was not associated with EPO-level in this population. CONCLUSIONS: In diabetic patients with CKD, elevated EPO-levels were predicted mainly by inflammatory markers and iron status, despite low hemoglobin levels and independent of kidney function. Therefore, anemia in diabetic CKD could be explained in part by inflammation-mediated iron dysregulation and resistance to endogenous EPO. KW - Diabetes mellitus KW - Anämie KW - Erythropoietin KW - Chronische Niereninsuffizienz KW - Entzündung KW - Inflammation KW - anemia KW - diabetes mellitus KW - chronic kidney disease KW - erythropoietin KW - inflammation Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-66637 ER - TY - JOUR A1 - Zanucco, Emanuele A1 - Götz, Rudolf A1 - Potapenko, Tamara A1 - Carraretto, Irene A1 - Ceteci, Semra A1 - Ceteci, Fatih A1 - Seeger, Werner A1 - Savai, Rajkumar A1 - Rapp, Ulf R. T1 - Expression of B-RAF V600E in Type II Pneumocytes Causes Abnormalities in Alveolar Formation, Airspace Enlargement and Tumor Formation in Mice JF - PLOS ONE N2 - Growth factor induced signaling cascades are key regulatory elements in tissue development, maintenance and regeneration. Perturbations of these cascades have severe consequences, leading to developmental disorders and neoplastic diseases. As a major function in signal transduction, activating mutations in RAF family kinases are the cause of human tumorigenesis, where B-RAF V600E has been identified as the prevalent mutant. In order to address the oncogenic function of B-RAF V600E, we have generated transgenic mice expressing the activated oncogene specifically in lung alveolar epithelial type II cells. Constitutive expression of B-RAF V600E caused abnormalities in alveolar epithelium formation that led to airspace enlargements. These lung lesions showed signs of tissue remodeling and were often associated with chronic inflammation and low incidence of lung tumors. The inflammatory cell infiltration did not precede the formation of the lung lesions but was rather accompanied with late tumor development. These data support a model where the continuous regenerative process initiated by oncogenic B-RAF-driven alveolar disruption provides a tumor-promoting environment associated with chronic inflammation. KW - obstructive pulmonary-disease KW - lung-cancer KW - somatic mutations KW - epithelial-cells KW - mouse models KW - protein KW - kinase KW - inflammation KW - activation KW - pathway Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-137061 VL - 6 IS - 12 ER - TY - THES A1 - Raab, Viktoria Maria T1 - Histologische Charakterisierung Vaccinia-Virus infizierter humaner Tumore im Mausmodell T1 - Histological charaterization of Vaccinia-Virus infected human tumors in mice N2 - Onkolytische Viren spielen eine immer bedeutendere Rolle für die Tumorforschung, weil in zahlreichen präklinischen Studien gezeigt werden konnte, dass viral bedingte Onkolyse zu einer Tumorregression führt. Ein äußerst vielversprechender Kandidat der onkolytischen Viren ist das Vaccinia-Virus. In der vorliegenden Arbeit wurde mit dem attenuierten Vaccinia-Virus GLV-1h68 gearbeitet, welches nach systemischer Applikation eine Regression von Tumoren verursacht. Obwohl bereits zahlreiche onkolytische Viren in klinischen Studien Anwendung finden, sind zugrunde liegende Abläufe bei einer Virusinfektion solider Tumore sowie Mechanismen, welche für die Tumorregression verantwortlich sind, immer noch nicht erschlossen. Um Aufschluss über notwendige Parameter für eine effiziente Infektion eines soliden Tumors mit GLV-1h68 zu erlangen, wurden im ersten Teil dieser Arbeit die uninfizierte Tumormikroumgebung sowie stromale Veränderungen in der frühe Phase der Infektion untersucht. Als Tumormodell diente hierbei ein humanes autologes Melanomzellpaar (888-MEL und 1936-MEL). Diese beiden Zelllinien sind Teil einer Reihe von fünf verschiedenen Melanomzelllinien, welche alle aus den widerkehrenden Metastasen eines einzelnen Patienten (Patient 888) isoliert wurden. 888-MEL zeigt nach Virusinfektion mit GLV-1h68 ein regredierendes Verhalten (therapeutischer Index: 88,0) und ist somit respondierend nach GLV-1h68-Infektion. 1936-MEL hingegen zeigte mit einem therapeutischen Index von 13,7 ein nur schwach verlangsamtes Wachstum solider Tumore, und ist somit schwach-respondierend nach GLV-1h68-Infektion. Als ein Grund, weshalb diese beiden autologen Melanomzelllinien unterschiedlich auf GLV-1h68-Infektion reagieren, wurde die Anzahl der Viruspartikel vermutet, welche 1 dpi im soliden Tumor vorliegt. Eine mögliche Korrelation zwischen initialem viralen Titer 1 dpi und späterer Tumorregression konnte experimentell aber nicht nachgewiesen werden. Zwei voneinander unabhängige Experimentreihen zeigten, dass bei identischer systemischer Applikation in den beiden soliden Tumoren kein Unterschied des viralen Titers vorlag. Weiterhin wurden die Komponenten der Tumormikroumgebung und ihr möglicher Einfluss auf die Effizienz der Virusinfektion untersucht. Immunhistologische Studien zeigten, dass es im uninfizierten Zustand bei soliden 888-MEL Tumoren zu einer massiven Infiltration CD45-positiver Zellen kam, die bei 1936-MEL-Tumoren jedoch nicht zu finden war. Die Beobachtung steht in Übereinstimmung mit Ergebnissen einer vergleichenden Microarray-Analyse, die das Infiltrat CD45-positiver Zellen in 888-MEL Tumoren genauer charakterisierte. Es wurde mit Microarray-Analyse eine erhöhte Expression chemotaktischer Moleküle in soliden 888-MEL Tumoren nachgewiesen. Unter anderem wird CCL8 (MCP-2) erhöht exprimiert. Als chemotaktisches Molekül hat CCL8 eine erhöhte Monozyteninfiltration zur Folge. Weiterhin wurde eine erhöhte Expression von MIF (macrophage migration inhibitory factor) und dem entsprechendem Rezeptor CD74 in uninfizierten 888-MEL-Tumoren gemessen. MIF induziert als proinflammatorisches Zytokin die Synthese inflammatorischer Mediatoren. Dies erklärt die Anhäufung CD45-positiver Zellen in der Tumormikroumgebung. Durch eine erhöhte Expression MHC II-verwandter Gene in soliden 888-MEL- Tumoren wurden die CD45-positiven Zellen als Monozyten identifiziert. Um die Funktion der Immunzellen zu analysieren, wurde durch eine intraperitoneale Applikation des Zytostatikums Cyclophosphamid eine Monozytendepletion induziert. Diese Immundepletion resultierte in soliden 888-MEL- Tumoren in einer signifikant verringerten Virusreplikation und -Ausbreitung nach Infektion mit GLV-1h68. Diese Ergebnisse implizieren, dass durch eine erhöhte Infiltration CD45-positiver Zellen in die Tumormikroumgebung die GLV-1h68-Infektion und -Replikation erleichtert wird. Nach Ausbreitung der Infektion kommt es in respondierenden Tumoren nach einem ersten Wachtumsarrest zu einer Tumorregression. Um Aufschluss über den beteiligten Mechanismus bei der Tumorregression zu erlangen, wurden GLV-1h68-infizierte-Tumore in der späten Phase der Infektion untersucht. Drei mögliche Mechanismen viral verursachter Onkolyse wurden beschrieben: Tumorzell-spezifische Onkolyse, Zerstörung der Tumorvaskulatur oder anti-tumorale Immunantwort. Für diese Experimente wurden humane Brustkarzinomzellen als Tumormodell verwendet. Mit diesem Tumormodell sollte analysiert werden, welcher der drei bislang diskutierten Mechanismen bei einer GLV-1h68-Infektion vorlag. Als erstes zeigten histologische Studien, dass Virusinfektion und -Replikation zu ausgedehnten Tumornekrosen führen. Dabei blieben die Blutgefäße in uninfizierten und auch in infizierten Bereichen des Tumors intakt und funktionell aktiv. Systemische Perfusion der Vaskulatur mit Lektin zeigte, dass die Tumorvaskulatur an das periphere Blutgefäßsystem angeschlossen war. Nachfolgende Experimente zeigten, dass Endothelzellen nicht durch die Viren infiziert wurden, wohingegen aber Endothelzell-ummantelnde, Gefäß-stabilisierende Perizyten nur in uninfizierten, nicht aber in infizierten Bereichen des Tumors vorkamen. Perizyten wurden möglicherweise durch Virusinfektion lysiert. Morphologische und funktionelle Analyse der Blutgefäße im Tumor zeigte, dass GLV-1h68-Infektion Hyperpermeabilität, Vasodilatation und eine erhöhte Expression des Adhäsionsmoleküls CD31 verursachte. Eine erhöhte CD31-Expression erleichtert eine Infiltration rekrutierter Immunzellen. Das konnte durch immunhistochemische Färbung von CD45 und MHC II besonders in intratumoralen Bereichen gezeigt werden. Durch Cyclophosphamid-vermittelte Immunsuppression wurde nachgewiesen, dass diese rekrutierten Immunzellen keinen ausschlaggebenden Einfluss auf die Tumorregression haben. Nach Immundepletion in soliden GI-101A-Tumoren konnte eine verstärkte Virusinfektion, effektivere Onkolyse und frühzeitigere Tumorregression nachgewiesen werden. Zusammenfassend zeigten diese Ergebnisse, dass der dominierende Mechanismus, welcher zur Tumorregression führt, die Onkolyse ist. N2 - Preclinical application of oncolytic viruses to induce virus-mediated tumor rejection revealed promising results in the past few years. Vaccinia virus GLV-1h68, used in this study, was shown to induce complete tumor disappearance upon infection. However the exact underlying mechanisms of viral infection and virus-mediated tumor regression are still not well understood. This study describes the characterization of the early phase of a GLV-1h68 infection and the role of stromal components as well as the mechanisms involved in tumor regression. To address the question, which components are necessary for an effective GLV-1h68 infection, followed by successive rounds of viral replication, the first part of this study focused on the characterization of the early phase of Vaccinia virus infection in a human autologous melanoma system. Five different cell lines were previously generated from a melanoma patient experiencing several reoccurrences in a 12 year span. The melanoma cell line expanded from a metastasis at an early time point (888-MEL) retained responsiveness to GLV-1h68 treatment, while the subsequent cell line 1936-MEL (studied here), became non-responsive to the same treatment. To address the influence of the amount of viral particles homing to the tumor within initial 24 hpi, 888-MEL and 1936-MEL tumors were compared. It could clearly be demonstrated, that the initial homing of viral particles is not the reason for further effective viral replication and spreading. The comparative viral titers were measured in 888-MEL and 1936-MEL tumors 1 dpi and found similar. Immuno-histological comparison of xenografts generated with 888-MEL or 1936-MEL revealed a massive infiltration of CD45-positive cells in 888-MEL tumors, but not in 1936-MEL tumors. Comparative microarray analysis of uninfected 888-MEL and 1936-MEL solid tumors supported these findings. Beside a significant up-regulation of CCL8 in 888-MEL tumors, an increased expression of CD74/MIF suggests an increased monocyte infiltration in 888-MEL uninfected tumors, which is not apparent in 1936-MEL tumors. To gain better understanding of an immune cell infiltration into solid 888-MEL tumors, a monocyte depletion study using the immunosuppressive agent cyclophosphamide (CPA) was carried out. This treatment resulted in a highly significant reduction of viral replication and spreading in 888-MEL tumors following infection with GLV-1h68. These results demonstrated that the replication efficiency of GLV-1h68 is higher in 888-MEL xenografts compared to 1936-MEL. The enhanced replication is in direct correlation with a higher number of CD45-positive cells, which infiltrated the tumor site prior to virus injection. Through successive rounds of viral replication the virus can spread throughout the tumor and induces tumor regression. To gain insights into mechanisms involved in tumor regression, late stages of a GLV-1h68 infection were characterized in human breast tumor xenografts (GI-101A). Theoretically oncolytic virus therapy could be based on three different mechanisms: by tumor cell specific oncolysis, by destruction of the tumor vasculature or by an anti-tumoral immunological response. Here the contribution of the three factors was analyzed. Histological examination showed that viral infection of GI-101A tumors led to broad tumor necrosis. However, the tumor vasculature in infected tumor areas remained functional and endothelial cells were not infected neither in tumors nor in cultured cells. It was further demonstrated, that viral tumor colonization activated the tumor endothelium leading to vascular hyperpermeability, vessel dilatation and an increased expression of the adhesion molecule CD31, which in a next step facilitated infiltration of inflammatory cell. This could be visualized by immunohistochemical staining of MHCII- and CD45-positive cells. The stainings revealed increased intratumoral infiltration of immune cells within infected tumor xenografts. The recruited immune cells however, do not seem to be causative for the tumorregression, since immunosuppression of GLV-1h68-infected animals led to increased viral replication and broader distribution in the tumor tissue, resulting in more efficient oncolysis and earlier start of the tumor regression phase. In summary, these results indicate that GLV-1h68 mediated oncolysis is the primary mechanism of tumor regression. Therefore, enhancing the viral replication and distribution within the tumor microenvironment should lead to improved therapeutic results in preclinical studies and clinical applications. KW - Vaccinia-Virus KW - Leukozyt KW - Allgemeine Entzündungsreaktion KW - Immunstimulation KW - Immunsuppression KW - Vaccinia-Virus KW - inflammation KW - leukocyte KW - innate immunity Y1 - 2010 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-49024 ER - TY - THES A1 - Grimm, Martin T1 - Die Bedeutung der Expression des Tumornekrosefaktor-alpha bei Patienten mit kolorektalem Karzinom T1 - Tumor Necrosis Factor-α expression in patients with colorectal cancer N2 - Für Tumorprogression müssen entartete Zellen Wege finden, die immunologische Abwehr und die Apoptose zu umgehen. Tumorzellen haben dafür verschiedene, sogenannte Tumor-Escape-Mechanismen entwickelt. Gegenstand dieser Arbeit war in diesem Zusammenhang die Untersuchung des TNF-α-TNF-R1-Systems. Eine signifikante erhöhte TNF-α Protein- und Genexpression konnte in Gewebeproben kolorektaler Karzinome nachgewiesen werden, wobei eine mäßig starke Korrelation beider Analysemethoden ersichtlich war (т = 0.794). Sowohl die immunhistochemische Analyse als auch die Genexpression durch RT-PCR konnten mit Tumorprogression assoziiert werden. Mit erhöhter Expression des Apoptose-induzierenden Zytokins TNF-α durch Tumorzellen konnte darüber hinaus ein signifikant schlechteres Gesamtüberleben der Patienten mit KRK beobachtet werden. In unmittelbarer Umgebung TNF-α exprimierender Tumorzellen wurden zahlreiche TNF-R1+/CD8+ Zellen analysiert, die als Hinweis auf Apoptose in TILs angesehen werden können. Dieser Weg könnte als Tumor-Escape-Mechanismus verstanden werden. Die Verwendung potentieller TNF-α Biologicals (z.B. Etanercept, Infliximab) bleibt jedoch unter dem Aspekt einer geringfügig erhöhten Lymphominzidenz gegenüber der Normalbevölkerung auch als kritisch zu bewerten. Der Einsatz von Anti-TNF-α-Therapien stellt jedoch eine vielversprechende Option bei Patienten mit metastasiertem KRK und Tumorrezidiv dar. Zusammenfassend liefern die Ergebnisse dieser Arbeit zusätzlichen Einblick in die Regulationsmechanismen, die verantwortlich für die Immunsuppression durch kolorektale Karzinome sein können. Diese basiswissenschaftlichen Erkenntnisse stellen eine mögliche Grundlage neuer Behandlungsmöglichkeiten kolorektaler Karzinomen dar, die weiter erforscht werden sollten. N2 - The progressive growth of malignancies is accompanied by a decline in the immune response through mechanisms which are poorly understood. Apoptosis and induction of inflammation by tumor released cytokines as tumor escape mechanisms have been proposed to play an important role in colorectal carcinogenesis. Expression of Tumor necrosis factor-alpha (TNF-α) was analyzed in colorectal cancer specimen by immunohistochemistry and RT-PCR. TNF-α expression on protein and mRNA level were correlated with clinical characteristics and impact on survival. TNFR-1 was co-labelled with TNF-α and CD8+ cytotoxic T cells in immunofluorescence double staining experiments. 94% of the patients with CRC expressed TNF-α. High TNF-α expression was significantly associated with positive lymph node stage and recurrence of the tumor. Multivariate analysis revealed high TNF-α expression as an independent prognostic factor. Immunohistochemistry was correlated with RT-PCR results (т = 0.794). Immunofluorescence double staining experiments revealed increased TNFR-1 expression by CD8+ cells. TNF-α expression by tumor cells may be an efficient immunological escape mechanism by inflammation-enhanced metastases and probably by induction of apoptosis in tumor-infiltrating CD8+ immune cells resulting in a down regulation of the tumoral immune response. Our data support the role of tumor-derived TNF-α expression as an important promoter of tumoral immune escape mechanisms and malignant progression. Targeting TNF-α (e.g. Etanercept, Infliximab) may be a promising option, especially in cases with high TNF-α expression and positive lymph node metastases. KW - Cancer KW - Tumor escape mechanism KW - death receptor signalling KW - apoptosis KW - inflammation KW - colorectal carcinoma Y1 - 2010 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-52757 ER - TY - THES A1 - Schwab, Nicholas T1 - The importance of CD8\(^+\) T cells and antigen-presenting cells in the immune reaction of primary inflammatory versus degenerative diseases T1 - Die Bedeutung CD8\(^+\) T-Zellen und Antigen-präsentierender Zellen in der Immunreaktion primär inflammatorischer gegenüber degenerativen Erkrankungen N2 - The bidirectional influence of parenchymal cells and cells of the immune system, especially of antigen-presenting and CD8\(^+\) T cells, in situations of putative auto- immune pathogenicity and degeneration was the main topic of this thesis. In the first part, the influence of human muscle cells on antigen-presenting cells was investigated. In inflammatory myopathies prominent infiltrates of immune cells containing T cells and antigen-presenting cells like macrophages and dendritic cells are present. The hypothesis was that human myoblasts have an inhibiting influence on these antigen-presenting cells under homeostatic conditions. A dysfunction or impairment under inflammatory circumstances might contribute to the development of myopathic conditions. The surface analysis of dendritic cells cocultured with myoblasts showed that immature dendritic cells could be driven into a reversible semi- mature state with significantly elevated levels of CD80. These dendritic cells were additionally characterized by their inhibiting function on T-cell proliferation. It was also shown that the lysates of healthy myoblasts could strongly enhance the phagocytic ability of macrophages, which could help with muscle regeneration and which might be disturbed in myositis patients. The second part of this thesis was about the clonal specificity of CD8\(^+\) T cells in a mouse model with genetically induced over-expression of PLP in oligodendrocytes. Here, we could show that the cytotoxic T lymphocytes, which had previously been shown to be pathogenic, were clonally expanded in the CNS of the transgenic mice. The amino acid sequences of the corresponding receptor chains were not identical, yet showed some similarities, which could mean that these clones recognize similar antigens (or epitopes of the same antigen). The knockout of PD-1 in this setting allowed for an analysis of the importance of tissue immune regulation. It became evident that the absence of PD-1 induced a larger number of clonal expansions in the CNS, hinting towards a reduced threshold for clonal disturbance and activation in these T cells. The expansions were, however, not pathogenic by themselves. Only in the presence of tissue damage and an antigenic stimulus (in our case the overexpression of PLP), the PD-1 limitation exacerbated the immune pathogenicity. Therefore, only in the presence of a “tissue damage signal”, the dyshomeostasis of T cells lacking PD-1 achieved high pathogenetic relevance. Finally, we investigated the pathogenetic role of CD8 T cells in Rasmussen encephalitis, a rare and chronic neurological disease mainly affecting children. The analysis of the T-cell receptor repertoire in Rasmussen encephalitis patients in the peripheral CD4\(^+\) and CD8\(^+\) T-cell compartments as well as the brain revealed the involvement of T cells in the pathogenicity of this disease. Many clonal expansions in the brain matched CD8\(^+\) T-cell expansions in the periphery on the sequence level. These putatively pathogenic clones could be visualized by immunohistochemistry in the brain and were found in close proximity to astrocytes and neurons. Additionally, the expanded clones could be found in the periphery of patients for at least one year. N2 - Der Einfluss von Parenchymzellen auf Immunzellen und umgekehrt, im Besonderen von Antigen-präsentierenden Zellen und CD8\(^+\) T-Zellen, im Zusammenhang von auto- immuner Pathogenese und Degeneration war das Hauptthema dieser Dissertation. Im ersten Teil wurde der Einfluss menschlicher Muskelzellen auf Antigen- präsentierende Zellen untersucht. In entzündlichen Myopathien kommt es zu massiven Infiltraten von Immunzellen, die T-Zellen und auch Antigen-präsentierende Zellen wie Makrophagen und dendritische Zellen enthalten. Die Hypothese war, dass menschliche Myoblasten einen hemmenden Einfluss auf die Antigen-präsentierenden Zellen unter homöostatischen Bedingungen haben. Eine Störung dieses Einflusses oder eine Beeinträchtigung unter entzündlichen Rahmenbedingungen könnte eventuell zur Entwicklung eines myopathischen Zustands beitragen. In der Oberflächenanalyse der dendritischen Zellen, die mit Myoblasten kultiviert wurden, zeigte sich, dass unreife dendritische Zellen in einen halb-reifen Zustand versetzt werden konnten, der sich beispielsweise durch stark erhöhte CD80 Expression kennzeichnet. Diese dendritischen Zellen wurden weiterhin charakterisiert über ihre hemmende Funktion auf die T-Zell Proliferation. Außerdem wurde gezeigt, dass Zelllysate gesunder Myoblasten die Phagozytoserate von Makrophagen enorm verstärken, was die Regeneration des Muskelgewebes erhöhen und möglicherweise in Myositispatienten gestört sein könnte. Im zweiten Teil der Dissertation ging es um die klonale Spezifität von CD8\(^+\) T-Zellen in einem Mausmodell mit genetisch induzierter Überexpression von PLP in Oligodendrozyten. Hier konnte gezeigt werden, dass die zytotoxischen T-Zellen, deren Pathogenität Gegenstand früherer Arbeiten war, im ZNS der transgenen Mäuse klonal expandiert waren. Die Aminosäuresequenzen der TCRβ Kette der expandierten Klone waren nicht identisch, zeigten jedoch einige Ähnlichkeiten, die darauf hinweisen könnten, dass diese Klone ähnliche Antigene (oder Epitope des gleichen Antigens) erkennen. Die genetisch induzierte Abwesenheit von PD-1 ermöglichte es, in diesem Zusammenhang den Einfluss von spezifischer Immunregulation im Gewebe zu untersuchen. Es zeigte sich, dass die Deletion von PD-1 eine erhöhte Anzahl von klonalen Expansionen im ZNS der Mäuse erzeugte, was auf eine herabgesetzte Schwelle für klonale Störungen und Aktivierung schließen lässt. Diese Expansionen 
 waren jedoch für sich genommen nicht pathogen. Nur in der Anwesenheit eines Gewebeschadens und eines zusätzlicher Antigenstimulus (in unserem Fall in Form der PLP Überexpression) konnte man die erhöhte Pathogenität durch die PD-1 Deletion erkennen. Deswegen erreichten die PD-1 deletierten T-Zellen nur in der Gegenwart eines „Gewebeschaden-Signals“ hohe pathogenetische Relevanz. Schließlich untersuchten wir die pathogenetische Rolle von CD8\(^+\) T-Zellen in der Rasmussen Enzephalitis, einer seltenen, chronischen Erkrankung des Gehirns, die hauptsächlich in Kindern vorkommt. Die Analyse des T-Zell-Rezeptor Repertoires in Rasmussen Enzephalitis Patienten in peripheren CD4\(^+\) und CD8\(^+\) T-Zell Populationen und im Gehirn zeigte die Beteiligung von T-Zellen in der Pathogenese dieser Krankheit auf. Viele klonale Expansionen waren zwischen Gehirn und der peripheren CD8\(^+\) Population bis hin zur Aminosäuresequenz identisch. Diese vermutlich pathogenen Klone konnten in Gehirnbiopsien von Rasmussenpatienten histochemisch nachgewiesen werden und wurden in enger Nachbarschaft zu Astrozyten und Neuronen gefunden. Zusätzlich konnten diese expandierten Kone in der Peripherie von Patienten für die beobachteten Zeiträume (mindestens ein Jahr) nachgewiesen werden. KW - T-Lymphozyt KW - Entzündung KW - Degeneration KW - Immunsystem KW - Rasmussen-Syndrom KW - T lymphocyte KW - inflammation KW - degeneration KW - immune system KW - rasmussen encephalitis Y1 - 2009 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-37330 ER - TY - THES A1 - Adamek, Anna Katharina T1 - Einfluss des Immunsystems und der endothelialen NO-Synthase auf den myokardialen ischämischen Schaden T1 - Influence of immune system and endothelial NO synthase on myocardial ischemic injury N2 - Die Entwicklung von therapeutischen Strategien, die den infarktbedingten Untergang des Myokardgewebes minimieren und die Gewebsheilung nach abgelaufenem Myokardinfarkt unterstützen, gehört zu dem Hauptziel in der modernen Kardiologie. Bis jedoch eine spezifische Intervention als Therapieform anerkannt wird, ist ein detailliertes Entschlüsseln der zellulären und molekularen Mechanismen während und nach der Myokardschädigung notwendig. Die vorliegende Arbeit beschäftigt sich intensiv mit den Vorgängen der Stickstoffmonoxid- (NO) Produktion und der Inflammation nach Okklusion von Kranzarterien. Im ersten Teil der Dissertation steht die endotheliale NO-Synthase-Expression (eNOS) im Mittelpunkt der Untersuchung. eNOS ist als wichtiger Katalysator an der Biosynthese von Stickstoffmonoxid, das als protektiver Faktor für die Gefäßhomöostase seit Jahren bekannt ist, beteiligt. Ferner besteht experimentell sehr gute Evidenz dafür, dass der endothelialen NO-Synthase am Ausmaß des kardialen Ischämie-/ Reperfusionsschadens eine entscheidende Rolle zukommt. Folglich wurde mittels der Substanz AVE 9488 versucht, die eNOS-Expression in Mäusen zu steigern und den Effekt auf das Infarktgeschehen näher zu betrachten. Die Behandlung mit AVE 9488 erzielte einen signifikant reduzierten Ischämie-/Reperfusionsschaden. Bei anschließenden Ischämie-/Reperfusionsveruchen mit eNOS defizienten Mäusen war der protektive Effekt wieder aufgehoben. Der Erfolg dieser Substanz wird in der signifikanten Reduktion des oxidativen Stresses vermutet. Ein zusätzlicher wichtiger Parameter, der während der Ischämie/Reperfusion aktiviert wird, ist der Schlüssel-Transkriptionsfaktor Nuclear Factor kappa B (NF-kB). Durch seine Bindung an bestimmte Enhancer und Promotoren reguliert der Faktor die Entzündungsprozesse, indem er die Genexpression proinflammatorischer Marker verstärkt. Folglich wurden eine Reduktion der Inflammation sowie ein protektiver Effekt nach erfolgter ischämischer Schädigung durch Hemmung von NF-kB angenommen. Zur Prüfung dieser Hypothese wurden NF-kB-Untereinheit p50 defiziente Mäuse (p50 KO) einer Okklusion einer Herzkranzarterie unterzogen. Durch die Hemmung der NF-kB-Aktivierung kam es zu einer signifikanten Reduzierung des Infarktareals im Vergleich zu den entsprechenden Wildtyp-Mäusen. Der große Benefit konnte auf die geringere Einwanderung der neutrophilen Granulozyten in das infarzierte Gebiet zurückgeführt werden. Knochenmarktransplantationsversuche mit p50 KO- und Wildtyp-Knochenmark untermauerten die Beobachtung, dass die beeinträchtigte Aktivierung von NF-kB in p50 defizienten Leukozyten protektive Effekte in der Ischämie/Reperfusion vermittelt. Die Aktivierung der proinflammatorischen Proteine während des linksventrikulären Remodelings nach Myokardinfarkt gehört zum Fokus des dritten Teils dieser Arbeit. Dieser Teil beschäftigt sich mit der Frage, inwieweit eine hochdosierte Aspirin-Therapie die linksventrikulären Umbauprozesse günstig beeinflussen kann. Dafür wurden Mäuse für 4 Wochen mit Placebo oder Aspirin (120 mg/kg pro Tag) mittels osmotischer Mini-Pumpen, die 2 Stunden nach Ligatur der Kranzarterie implantiert wurden, behandelt. In beiden Gruppen kam es zur erwarteten linksventrikulären Dilatation nach Myokardinfarkt, jedoch ohne signifikanten Unterschied zwischen Placebo- und Aspirin-behandelten Tieren. Es kam allerdings zu einer erwarteten Reduktion proinflammatorischer Proteine durch die Aspirin-Therapie. So war die Expression von Tumor-Nekrose-Faktor-alpha; (TNF-alpha) und Interleukin-1ß (IL-ß) in der Aspirin-Gruppe signifikant reduziert. Zusammenfassend lässt sich sagen, dass durch die gezielte Beeinflussung bestimmter Faktoren in der Ischämie/Reperfusion wie z. B. die Verstärkung der eNOS-Expression oder die Hemmung der NF-kB-Aktivierung die Ischämieschädigung signifikant reduziert werden kann. N2 - One of the major therapeutic goals of modern cardiology is to design strategies aimed at minimizing myocardial necrosis and optimizing cardiac repair following myocardial infarction. However, a sound understanding of the cellular and molecular mechanism is necessary before a specific intervention is pursued on a therapeutic basis. The present work includes important aspects of inflammation and nitric oxide (NO) production after occlusion of the coronary artery. The first part of the thesis focused on endothelial NO synthase (eNOS). eNOS is a promotor of NO biosynthesis, which regulates vascular and myocardial function. Moreover, endothelial NOS is cardioprotective in ischemia/reperfusion injury. Therefore, the effects of AVE 9488, a novel pharmacon designed to selectively increase eNOS protein expression and NO formation, was tested on cardiac ischemia/reperfusion injury in vivo. Ischemia/reperfusion damage was significantly reduced in mice treated with AVE 9488 when compared to placebo treated mice. The protective effect was blunted in eNOS knockout mice treated with the eNOS enhancer. The expression of inflammatory markers was not influenced by the therapy. Reactive oxygen species were significantly reduced in mice treated with the eNOS enhancer. In addition, the transcription factor nuclear factor kappa B (NF-kB) is important in cardiac damage. NF-kB is activated by various stimuli implicated in ischemia/reperfusion injury and increases the expression of proinflammatory markers by binding on special enhancer and promoters. Inhibition of NF-kB might therefore reduce the inflammatory response and achieve protective effects after myocardial infarction. To prove this hypothesis mice with targeted deletion of the NF-kB subunit p50 (p50 KO) underwent 30 minutes of coronary artery ligation and 24 hours of reperfusion in vivo. Ischemia/reperfusion damage was significantly reduced in the p50 KO animals when compared with matching wild-type (WT) mice. Although adhesion molecules such as intercellular adhesion molecule were up-regulated in left ventricles of p50 KO mice, fewer neutrophils infiltrated the infarct area, suggesting leukocytes as a potential mediator of the protection observed in the p50 KO. This was confirmed in adoptive transfer experiments: whereas transplantation of KO bone marrow in KO animals sustained the protective effect on ischemia/reperfusion injury, transplantation of WT bone marrow in KO animals abolished it. The last part tested the hypothesis that inhibition of the proinflammatory response to myocardial infarction could improve left ventricular remodeling. Therefore, mice were treated for 4 weeks with placebo or aspirin (120 mg/kg per day) by Alzet mini-osmotic pumps implanted 2 hours after ligation of the left anterior descending artery. On echocardiography, animals 4 weeks after myocardial infarction exhibited left ventricular dilatation as expected. However, there was no difference between the placebo and the Aspirin group. The expression of the proinflammatory cytokines tumor necrosis factor alpha; (TNF-alpha) and interleukin 1ß (IL-1ß) which were markedly upregulated in mice with myocardial infarction on placebo were significantly reduced by Aspirin treatment. However, left ventricular remodeling after myocardial infarction was not altered. In conclusion, the use of specific strategies to inhibit the NF-kB activation or to increase eNOS expression in ischemia/reperfusion constitutes a promising novel therapeutic approach to reduce ischemic damage. However, successful application of anti-inflammatory interventions in the treatment of ischemic remodeling will require a better understanding of the specific molecular steps in the regulation of cardiac injury and repair. KW - Ischämie KW - Reperfusion KW - Entzündung KW - Oxidativer Stress KW - ischemia KW - reperfusion KW - inflammation Y1 - 2008 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-35795 ER - TY - THES A1 - Müller, Verena T1 - Candida albicans-induzierte Genexpression in primären humanen Endothelzellen - Mechanismen der Signaltransduktion und Möglichkeiten der Intervention T1 - Candida albicans-induced gene expression in primary human endothelial cells - mechanisms of signal transduction and possibilities of intervention N2 - Endothelzellen sind ein aktiver Bestandteil der angeborenen Immunabwehr des Menschen gegen mikrobielle Pathogene. Unter ungünstigen Bedingungen kann die Abwehrreaktion sogar zu einer lebensbedrohlichen Sepsis führen. Hier wurde die bislang wenig bekannte Endothelantwort auf den fakultativ humanpathogenen Hefepilz Candida albicans, einem der häufigsten Verursacher von letaler Sepsis beim Menschen, näher untersucht. Mittels Oligonukleotid-Mikroarray-Analyse von HUVEC nach Exposition mit C. albicans konnten 56 hochregulierte Gene identifiziert werden, während 69 Gene herunterreguliert wurden. Ein bedeutender Anteil der regulierten Gene ist an Prozessen der angeborenen Immunantwort beteiligt und dient hauptsächlich der Rekrutierung von Neutrophilen. Weitere Untersuchungen ergaben eine zentrale Rolle des proinflammatorischen NF-kappaB-Weges bei der Regulation des Candida-induzierten Transkriptoms von Endothelzellen. Es konnte gezeigt werden, dass C. albicans diesen Signalweg sequenziell aktiviert. Zusätzlich konnte durch die Expression einer dominant-negativen Mutante einer Signalkomponente des NF-kappaB-Signalwegs die Candida-vermittelte Induktion von kappaB-abhängigen Genen gehemmt werden. Mit einem pharmakologischen Ansatz wurde der p38 MAP Kinase-Signalweg als weiterer bedeutsamer Signalweg identifiziert, der die Expression einzelner Candida-Zielgene wie CXCL8/IL-8 moduliert. Schließlich wurde gezeigt, dass die Candida-induzierte NF-kappaB-Aktivierung im untersuchten endothelialen Zellsystem unabhängig von den Toll-like Rezeptoren TLR2 und TLR4 geschieht, die üblicherweise an der Erkennung mikrobieller Pathogene beteiligt sind. Durch RNA-Interferenz-Experimente konnte jedoch dargelegt werden, dass das Adaptermolekül MyD88 und die Kinase IRAK1, die beide entscheidend an der TLR-vermittelten Signaltransduktion beteiligt sind, essentiell für die Weiterleitung des Signals in Endothelzellen sind. Nachfolgend konnte mit TLR3 zumindest einer der signaltransduzierenden Rezeptoren identifiziert werden. Als erste umfassende Untersuchung der endothelialen Antwort auf Candida albicans erlaubt die vorliegende Arbeit neue Einblicke in die komplexen Signalmuster von Endothelzellen, die dieser klinisch bedeutende Krankheitserreger auslöst. N2 - Endothelial cells (ECs) actively participate in the innate defence against microbial pathogens. Under unfavourable conditions defence reactions can even turn into life-threatening responses resulting in sepsis. Here the so far largely unknown EC reaction patterns to Candida albicans were studied. C. albicans is a facultative human pathogenic fungus and a major cause of lethality in septic patients. Oligonucleotide microarray analysis revealed 56 genes that were transcriptionally up-regulated and 69 that were suppressed upon exposure of ECs to C. albicans. A major portion of these genes is involved in defence mechanisms of the innate immune system with a high representation of genes serving the recruitment of neutrophils to sites of infection. Further examination of candidate signalling cascades established a central role of the proinflammatory NF-kappaB pathway in the regulation of the Candida-modulated transcriptome of ECs. It was shown that the NF-kappaB signalling pathway becomes activated at various levels. In addition, expression of a dominant negative mutant of a NF-kappaB signalling pathway compound blocked the Candida-induced kappaB-dependent gene expression. Using a pharmacological approach the stress-activated p38 mitogen-activated protein (MAP) kinase pathway was identified as a second major regulatory pathway which critically contributes to the regulation of selected Candida target genes such as CXCL8/IL-8. Candida-induced NF-kappaB activation is mediated independently of Toll-like receptors (TLR) 2 and 4 that commonly have been implicated with microbial pattern recognition. Nevertheless, knock-down of the adapter molecule MyD88 and the essential downstream kinase of TLR-, IL-1R- and IL-18R-signalling, IRAK1, suggested that recognition and signalling via a TLR apart from TLR2/TLR4 is crucial for Candida-induced gene expression in primary ECs. Finally, RNAi experiments indicate that most likely TLR3 represents this receptor. These data provide the first comprehensive analysis of endothelial gene responses to Candida albicans and present novel insights into the complex signalling patterns triggered by this important pathogen. KW - Angeborene Immunität KW - Endothelzelle KW - Toll-like-Rezeptoren KW - Candida albicans KW - Entzündung KW - innate immunity KW - endothelial cell KW - toll-like receptors KW - Candida albicans KW - inflammation Y1 - 2007 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-26224 ER - TY - THES A1 - Schopf, Thilo T1 - Wertigkeit von Entzündungsparametern in der postoperativen Phase nach Implantation von Hüft- und Kniegelenk-Totalendoprothesen T1 - relevance of inflammatory parameters after hip and knee arthroplasty N2 - Ziel dieser Arbeit ist, die Wertigkeit von Entzündungsparametern (CRP, BSG, Leukozyten und Temperatur) in der postoperativen Phase anhand einer retrospektiven Studie mit 531 Patienten nach Implantation von Knie- und Hüft-Totalendoprothesen zu beurteilen. Hierbei sollten zum einen die Auswirkungen von Vorerkrankungen (Asthma bronchiale, Rheumatoider Arthritis, Nikotinabusus, Gicht/Hyperurikämie, Z. n. Thrombosen) auf den postoperativen Verlauf der Entzündungsparameter und zum anderen postoperative Komplikationen (Thrombosen, bronchopulmonale Infekte, Harnwegsinfekte, Wundinfekte, Protheseninfekte) anhand der Entzündungsparameter erfasst werden. Die Auswertung erfolgte mittels SPSS für Windows durch den Mann-Whitney Test als nicht parametrischen Test für unabhängige Variablen. Im Durchschnitt stieg der CRP-Wert von 0,6 mg/dl präoperativ auf etwa 8,5 mg/dl um den 3. postoperativen Tag an und fiel dann durchschnittlich auf 1,4 mg/dl um den 14. postoperativen Tag ab. Die BSG-Werte stiegen von etwa 18 mm/h präoperativ auf 52 mm/h um den 3. postoperativen Tag an. Der höchste BSG-Wert (56 mm/h) wurde erst um den 7. postoperativen Tag erreicht. Danach fiel der BSG-Wert leicht ab auf durchschnittlich etwa 43 mm/h um den 14. postoperativen Tag. Die Temperaturkurve verlief ähnlich: Auch hier stieg die Temperatur rasch auf einen Höchstwert von 37,1°C um den 2. postoperativen Tag. Es folgte ein langsamer Abfall der Temperatur, bis hin zu Normwerten ab dem 10. postoperativen Tag. Die Leukozyten zeigten im Durchschnitt einen geradlinigen Verlauf mit durchschnittlichen Werten zwischen 7 und 8 G/l, allerdings bei einer großen Standartabweichung. Patienten mit Asthma bronchiale zeigten präoperativ signifikant erhöhte BSG und Leukozytenwerte. Patienten mit Gicht/Hyperurikämie fielen durch signifikant niedrigere Temperaturwerte am 6. und 12. postoperativen Tag sowie durch Trends zu niedrigeren Werten am 10. und 16. postoperativen Tag auf. Signifikant höhere Werte fanden sich bei der CRP-AUC-Kurve vom 7.-14. postoperativen Tag. Des Weiteren fanden sich Trends zu höheren Werten am 14. postoperativen Tag bei der BSG und am 3. postoperativen Tag bei den Leukozyten. Bei Patienten mit Z.n. Thrombose zeigte sich der postoperative BSG-Verlauf vom 3.-14. Tag signifikant erniedrigt, ebenso waren die postoperativen Temperaturwerte vom 15. und 16. Tag signifikant niedriger. Der einzig signifikante Unterschied zwischen Rauchern und Nichtrauchern fand sich in der präoperativen BSG, wo Raucher signifikant niedrigere Werte hatten. Des Weiteren zeigten sich Trends zu höheren Leukozytenwerten am 3. und 7. postoperativen Tag sowie Trends zu niedrigeren Temperaturen am 1., 2., 7., 8. und 14. postoperativen Tag. Patienten mit Rheumatoider Arthritis fielen durch signifikant erhöhte präoperative CRP- und BSG-Werte auf. Zusätzlich traten Trends zu höheren Werten am 14. postoperativen Tag für CRP, BSG und Leukozyten auf. Dem Trend zur Temperaturerhöhung am 2. postoperativen Tag folgten erneute Trends am 9. und 11. postoperativen Tag sowie signifikant höhere Temperaturen am 10. und 12. postoperativen Tag. Patienten, die einen Harnwegsinfekt erlitten, hatten signifikant erhöhte Werte am 14. postoperativen Tag für CRP und BSG. Auch die CRP-AUC-Kurve zeigte diesen Verlauf signifikant. Bei Patienten, die an einem bronchopulmonalem Infekt erkrankten, war der CRP-Wert des 3. postoperativen Tages signifikant erhöht. Dies ließ sich auch in der CRP-AUC-Kurve vom 0.-3. postoperativen Tag signifikant nachweisen. Darüber hinaus lag ein Trend am 3.-7. postoperativen Tag zu höheren Werten vor. Bei Patienten mit tiefer Beinvenenthrombose fiel eine signifikante Temperaturerhöhung am 7., 8. und 11. postoperativen Tag auf, sowie ein Trend am 9., 10. und 12. postoperativen Tag. Die CRP-Werte stiegen um den 14. postoperativen Tag signifikant an. Gleiches Verhalten zeigte die CRP-AUC-Kurve vom 7.-14. postoperativen Tag. Patienten mit Wundinfektionen fielen durch Trends zu niedrigeren Temperaturwerten am 4. und 5. postoperativen Tag auf. Außerdem stieg um den 7. postoperativen Tag die BSG trendwertig an. Die Leukozyten waren präoperativ signifikant erhöht. Dagegen war bei Patienten mit TEP-Infektion der CRP-Wert um den 14. postoperativen Tag signifikant erhöht. Die Temperaturwerte des 8. postoperativen Tages zeigten einen Trend zu niedrigeren Zahlen. Die Ergebnisse entsprechen in etwa denen anderer Studien. Insgesamt lässt sich eine Tendenz ablesen: BSG und Temperatur sind vor allem bei den erwähnten Vorerkrankungen erhöht, Leukozyten spielen kaum eine Rolle und die CRP-Bestimmung findet ihren Platz hauptsächlich in der Diagnostik von Komplikationen. Außerdem wird klar, dass kein Entzündungsparameter spezifisch für eine bestimmte Vorerkrankung oder Komplikation ist. Darüber hinaus sind nicht Einzelwerte, sondern viel mehr der Verlauf der Entzündungsparameter entscheidend. N2 - Blood parameter like CRP (C-reactive protein), ESR (erythrocyte sedimentation rate), white blood cells (leukocytes) and body temperature were messured after total hip and knee arthroplasty in 531 patients. The effects of bronchial asthma, rheumatiod arthritis, smoking, gout and a deep vein thrombosis in the past on inflammatory parameters were investigated, on the oher side we tried to check out, whether deep vene thrombosis, bronchial or pulmonary infections, urinary tract infections, wound infections or infections of the endoprothesis influence the developing of inflammatory parameters. KW - Totalendoprothese KW - Entzündung KW - Hüftgelenkentzündung KW - TEP KW - Komplikation KW - Laborparameter KW - Entzündungsparameter KW - postoperative Komplikationen KW - Endoprothese KW - inflammation KW - hip KW - knee KW - arthroplasty KW - complications Y1 - 2007 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-25028 ER - TY - THES A1 - Wienrich, Bernd Gregor T1 - Expression von LEEP-CAM (Lymphocyte Endothelial EPithelial-Cell Adhesion Molecule) in Haut und Hoden - funktionelle Implikationen für die Immunevasion epithelialer Hauttumoren und Entzündungen immunprivilegierter Gewebe T1 - Expression of LEEP-CAM (Lymphocyte Endpthelial EPithelial-Cell Adhesion Molecule) in Skin and testis –functional implication of immunevasion in epithelial skincancer and inflammation in immunologically privileged tissue N2 - Der Schutz vor der Einwanderung von Immunzellen ist einerseits unter physiologischen Bedingungen wichtig für die Integrität immunprivilegierter Organe, andererseits aber auch (mit)entscheidend für die Pathogenese maligner Tumoren. Vor diesem Hintergrund wurde LEEP-CAM (Lymphocyte Endothelial EPithelial-Cell Adhesion Molecule) untersucht, ein Adhäsionsmolekül, welches in der Epidermis und den dermalen Blutgefäßen in normaler Haut konstitutiv exprimiert wird. Durch immunhistochemische Untersuchungen wurde im ersten Teil der Arbeit gezeigt, dass LEEP-CAM in Basalzellkarzinomen, Plattenepithelkarzinomen und Keratoakanthomen der Haut deutlich vermindert oder gar nicht exprimiert wird. Die verminderte Expression war mit fehlender Infiltration von T-Lymphozyten in das Tumorgewebe assoziiert, was insbesondere durch zwei hinsichtlich ihrer LEEP-CAM-Expression unterschiedenen Populationen von Keratoakanthomen nahe gelegt wurde. Die Hypothese, dass LEEP-CAM in die epidermale Rekrutierung aktivierter T-Zellen involviert ist, wurde durch funktionelle Stamper-Woodruff-Experimente (Adhäsion aktivierter T-Lymphozyten an Gewebe-Gefrierschnitte) mit Basalzellkarzinomen und psoriatischer Haut gestützt. Durch metabolische Markierung mit 35(S)-Methionin und anschließende Radioimmunpräzipitation sowie durch durchflusszytometrische Untersuchungen an kultivierten Zellen wurde gezeigt, dass LEEPCAM in transformierten Keratinozyten im Vergleich zu normalen Keratinozyten deutlich vermindert synthetisiert und exprimiert wird. In zwei komplementären murinen Karzinogenese-Modellen wurde die Assoziation der verminderten LEEP-CAM-Expression mit Entdifferenzierung und invasivem Wachstum der Tumorzellen untermauert. Insgesamt kann experimentelle Evidenz für die Hypothese, dass die Herabregulation der LEEP-CAM-Expression ein (Teil)-Mechanismus ist, durch welchen sich invasiv wachsende Tumoren den Angriffen des Immunsystems entziehen können, präsentiert werden. Im Weiteren wurde die Expression und Funktion von LEEPCAM im Keimepithel des Hodens (als ein Beispiel für ein immunprivilegiertes Gewebe) untersucht. Durch immunhistochemische Untersuchungen wurde die konstitutive Expression von LEEP-CAM in den Sertoli-Zellen des Keimepithels nachgewiesen. Mittels Immun-Elektronenmikroskopie wurde dann die Lokalisation an desmosomalen Strukturen sowie entlang der Zellmembran gezeigt. Im Hinblick auf die Funktion von LEEP-CAM wurde in modifizierten Stamper-Woodruff-Experimenten erstmals gezeigt, dass aktivierte T-Lymphozyten an das Keimepithel des Hodens binden können und dass diese Adhäsion durch LEEP-CAM-gerichtete Antikörper inhibiert werden kann. Damit ist LEEP-CAM das erste Molekül, für welches direkte experimentelle Evidenz eine mögliche Rolle bei der testikulären Lymphozyten-Rekrutierung belegt. Dies könnte Relevanz für die Pathogenese von Orchitiden, den häufigsten Ursachen männlicher Infertilität, haben. N2 - Under physiologic conditions protection of invading immune cells is on one hand important for integrity of immune-privileged organs and on the other hand crucial for the pathogenesis of malignant tumors. In the current thesis LEEP-CAM (Lymphocyte Endothelial Epithelial-Cell Adhesion Molecule), an adhesion molecule which is expressed in the epidermis and dermal blood vessels was investigated. In immunohistochemical stains LEEP-CAM expression was dramatically reduced or absent in basal cell carcinoma, squamous cell carcinoma or keratoacanthoma. Reduced expression was associated with lack of infiltrating T-lymphocytes in the tumor tissue, which was especially obvious in keratoacanthoma. The hypothesis that LEEP-CAM is involved in epidermal recruiting of activated T cells was verified by functional Stamper-Woodruff experiments (adhesion of activated T lymphocytes to frozen tissue sections) for basal cell carcinoma and psoriasis. By metabolic labeling with 35-S-methionin and subsequent radioimmuno-precipitation and additional FACS analysis we could demonstrate, that LEEP-CAM synthesis and expression was reduced in transformed keratinocytes in comparison to primary keratinocytes. The association of reduced LEEP-CAM expression with differentiation and invasive tumor growth was confirmed in two complementary murine carcinogenesis models. In summary, reduced expression of LEEP-CAM may be a mechanism by which invasive tumors are capable of escaping the immune surveillance. In the second part of the thesis, expression and function of LEEP-CAM in the testis was investigated. Constitutive expression of LEEP-CAM was identified immunhistochemically in Sertoli cells. By immunoelectron microscopy LEEP-CAM was localized to desmosomal structures and along the cell membrane. Functional analysis by modified Stamper-Woodruff experiments demonstrated the capability of activated T-lymphocytes to bind to germinal tissue of human testis. This adhesion could be blocked by anti-LEEP-CAM-antibodies. Our studies suggest that LEEP-CAM may the first molecule which is involved in lymphocyte testicular recruitment. This may be relevant for the pathogenesis of orchtitis. KW - Zell-Adhesionsmolekül KW - Hoden KW - Haut KW - Hauttumor KW - Hautkrebs KW - Basaliom KW - Epitheliom KW - Spinaliom KW - Endothel KW - Epithel KW - Keimepithel KW - Plattenepithel KW - Keratoakant KW - LEEP-CAM KW - adhesion molecule KW - inflammation KW - human testis KW - LEEP-CAM KW - adhesion molecule KW - inflammation KW - human testis Y1 - 2006 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-27872 ER - TY - THES A1 - Timmermann, Meike T1 - Zelluläre Regulation und klinische Aspekte des monocyten-/macrophagenspezifischen Proteins CD163 T1 - Cellular regulation and clinical aspects of the monocyte/macrophage-specific protein CD163 N2 - In der vorliegenden Arbeit wurde die zelluläre Regulation des monocyten-/ macrophagenspezifischen Oberflächenproteins CD163 untersucht und klinische Aspekte der löslichen Form des CD163 (sCD163) diskutiert. sCD163 wird in vivo durch einen inflammatorischen Reiz von der Zelloberfläche abgespalten. Bislang waren jedoch noch keine Mediatoren charakterisiert worden, die immer unter Entzündungsbedingungen vorhanden sind. In den eigenen Untersuchungen des Shedding von CD163 konnten für die Generierung des sCD163 neue endogene Aktivatoren identifiziert werden. Sowohl reaktive Sauerstoffspezies, wie Wasserstoffperoxid oder Stickstoffmonoxid, als auch das Produkt von endogenen Oxidationsreaktionen mit reaktiven Sauerstoffspezies 8-iso Prostaglandin F2a erwiesen sich als potente Aktivatoren des Shedding von CD163. Neben den bekannten physiologischen Funktionen des 8-iso Prostaglandin F2a konnte erstmals eine neue Funktion bei Entzündungen definiert werden. Dieser Effekt wurde spezifisch durch 8-iso Prostaglandin F2a hervorgerufen, da das isomere Prostaglandin F2a unter gleichen Bedingungen keinen Einfluss auf das Shedding von CD163 ausübte. Das Immunsuppressivum Cyclosporin A konnte ebenfalls als Induktor des Shedding von CD163 ermittelt werden. Damit konnte zusätzlich zur bekannten immunmodulatorischen Wirkung des Cyclosporin A eine weitere antiinflammatorische Wirkung über Monocyten/Macrophagen aufgezeigt werden. Durch Untersuchungen der Inhibierung des Shedding von CD163 konnten Gemeinsamkeiten bezüglich der in die sCD163-Generierung involvierten Mediatoren dargestellt werden. Obwohl die untersuchten Verbindungen wahrscheinlich über unterschiedliche Signalwege das Shedding von CD163 induzieren, waren die Anwesenheit von reaktiven Sauerstoffspezies und intrazellulärem Calcium und die Beteiligung einer TIMP-3-sensitiven Metalloproteinase an diesem Prozess essentiell. Bei einem Vergleich zwischen dem Shedding von CD163 und Tumor necrosis factor-a (TNF-a) ergaben sich Gemeinsamkeiten durch die Induktion des Shedding beider Verbindungen durch 8-iso Prostaglandin F2a und in sehr viel geringerem Maße durch Wasserstoffperoxid. Im Gegensatz dazu waren deutliche Unterschiede in dem Ausmaß der induzierten Stimulation des Shedding durch Stickstoffmonoxid, Prostaglandin F2a und Cyclosporin A zu erkennen. Im Hinblick auf die entgegengesetzten Wirkungen von sCD163 als antiinflammatorisch wirkende Verbindung und TNF-a als proinflammatorisches Cytokin, konnte dargestellt werden, dass die Freisetzung durch unterschiedliche Aktivatoren erfolgt. Nach Bestimmung der Konzentrationen von sCD163 und 8-iso PGF2a in bronchoalveolärer Lavage-Flüssigkeit von Patienten mit Cystischer Fibrose konnten keine abschließende Aussagen über die Eignung beider Parameter als biologische Marker für chronische Entzündungen der Lunge bei diesen Patienten getroffen werden. Weiterführend zu der Kenntnis, dass sCD163 die antiinflammatorische Wirkung über eine Interaktion des Proteins mit humanen T-Lymphocyten und nachfolgender Hemmung der Proliferation dieser Zellen ausübt, wurde diese Wechselwirkung genauer untersucht. In quantitativen Bestimmungen des sCD163 in isolierten T-Lymphocyten verschiedener Spender konnte erstmals gezeigt werden, dass sCD163 zu einem Teil konstitutiv in die T-Lymphocyten aufgenommen wird und dass diese Aufnahme durch proinflammatorische Aktivierung der T-Lymphocyten stark gesteigert werden kann. Durch fluoreszenzmikroskopische Aufnahmen unstimulierter T-Lymphocyten konnte die intrazelluläre Lokalisation des sCD163 visualisiert werden. Nach Aktivierung der Zellen mit einem proinflammatorischen Reiz fand innerhalb der Zellen eine Translokalisation des sCD163 aus dem cytoplasmatischen Bereich zur Zellmembran statt. Damit konnte erstmals gezeigt werden, dass abhängig vom Aktivierungsstatus der T-Lymphocyten eine Umverteilung des sCD163 innerhalb der Zellen erfolgt. Eine quantitative Bestimmung des sCD163 und seines Bindungspartners in T-Lymphocyten nichtmuskuläres Myosin Typ IIA gelang mittels eines neu entwickeltem ELISA, der spezifisch sCD163 und Myosin ausschließlich im Komplex erfasst. Damit konnte durch die in dieser Arbeit beschriebenen Untersuchungen ein grundlegender Beitrag zur Charakterisierung der Regulation der Proteinexpression und des Shedding von CD163 in humanen Monocyten sowie der Interaktion des sCD163 mit T-Lymphocyten geleistet werden. N2 - In the present thesis, cellular regulation of the monocyte/macrophage specific membrane protein CD163 was investigated and clinical aspects of the soluble form of CD163 (sCD163) were discussed. sCD163 is shed from the cell surface in vivo upon an inflammatory stimulus. So far no mediators had been characterized that are persistently present under inflammatory conditions. In the present investigation of the shedding of CD163, new endogenous activators for the generation of sCD163 were successfully identified. Both reactive oxygen species, as hydrogen peroxide or nitric monoxide, and the product of endogenous oxidative reactions 8-iso prostaglandin F2a turned out to be potent activators of the shedding of CD163. In addition to the known physiological functions of 8-iso prostaglandin F2a a novel function of this compound in inflammation was defined. This effect was specifically generated by 8-iso prostaglandin F2a as prostaglandin F2a did not influence the shedding of CD163 under the same conditions. The immunosuppressive drug cyclosporine A was also established as an inductor of the shedding of CD163. Accordingly, another anti-inflammatory effect via monocytes/macrophages was pointed out in addition to the well known immunomodulatory effects of cyclosporine A. Investigations of the inhibition of shedding of CD163 led to the identification of key mediators involved in the generation of sCD163. Even though the tested compounds may induce shedding via different signal transduction pathways, the presence of reactive oxygen species and intracellular calcium and the involvement of a TIMP-3 sensitive metalloproteinase were essential in this process. The tested activators revealed different abilities for inducing the formation of reactive oxygen species. Comparing shedding of CD163 and tumor necrosis factor-a (TNF-a), similarities in induced shedding by 8-iso prostaglandin F2a and to a lower extend by hydrogen peroxide were seen. In contrast, significant differences were recognized in the extent of shedding induced via stimulation by nitric oxide, prostaglandin F2a, and cyclosporine A. With regard to the opposite effects of sCD163 as an anti-inflammatory compound and TNF-a as a pro-inflammatory cytokine it was demonstrated that shedding occurred via induction by different activators. The determination of concentrations of sCD163 and 8-iso prostaglandin F2a in bronchoalveolar lavage fluid of patients with cystic fibrosis allowed no conclusive statement about the suitability of both parameters as marker for chronic inflammation. In extension to earlier insights in the anti-inflammatory effects of sCD163 via interaction of the protein with human T-lymphocytes and subsequent inhibition of the proliferation of these cells these interactions were analyzed in detail. Quantifications of sCD163 in isolated T-lymphocytes from different donors revealed for the first time that sCD163 is constitutively taken up into T-lymphocytes and that this process can significantly be enhanced by pro-inflammatory activation of T-lymphocytes. The intracellular localization of sCD163 was visualized by fluorescence microscopy of T-lymphocytes. Upon activation of the cells by a pro-inflammatory stimulus a translocalization of sCD163 was observed within the cells from the cytoplasmatic region to the cell membrane. Thus, it was shown for the first time that an activation dependent translocalization of sCD163 occurs within the T-lymphocytes. The quantitative determination of sCD163 and non-muscular myosin type IIA as its intracellular binding partner in T-lymphocytes was successful using a newly developed ELISA that detects specifically sCD163 and myosin as a complex. To conclude, the investigations described in this thesis contributed substantially to the understanding of the regulation of the protein expression and shedding of CD163 in human monocytes and of the interaction of sCD163 with T-lymphocytes. KW - Antigen CD163 KW - Monozyt KW - Entzündung KW - Prostaglandine KW - CD163 KW - Monocyten KW - Regulation KW - Entzündung KW - Isoprostaglandin KW - CD163 KW - monocytes KW - regulation KW - inflammation KW - isoprostaglandin Y1 - 2005 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-16028 ER -