TY - THES A1 - Westfahl, Larissa T1 - Influenza-assoziierte Hospitalisierungen bei Kindern und Erwachsenen am Universitätsklinikum Würzburg in den Jahren 2010-2013 T1 - Influenza-associated hospitalizations at children and adults at University hospital Würzburg in 2010-2013 N2 - Influenza-assoziierte Atemwegserkrankungen führen jedes Jahr zu zahlreichen Hospitalisierungen und Todesfällen. Der während der Pandemie 2009 zirkulierende Erreger Influenza A(H1N1)pdm09 führte zu zahlreichen, zum Teil schweren Komplikationen, insbesondere auch bei jüngeren Erwachsenen. In der vorliegenden Studie wurden Influenza-assoziierte Hospitalisierungen (IAH) hinsichtlich Krankheitsverlauf in den verschiedenen Altersgruppen sowie bei verschiedenen Erregern untersucht. Zudem erfolgte eine Analyse der direkten Krankheitskosten. Einen besonders schweren Verlauf zeigten Erwachsene mit Grunderkrankung zwischen 18-60 Jahren, die überwiegend mit dem Erreger Influenza A(H1N1)pdm09 infiziert waren. Ebenso waren schwangeren Patientinnen mit IAH mit dem Erreger A(H1N1)pdm09 selten, aber schwer betroffen. Bei Patienten von 18-60 Jahren mit dem Erreger Influenza A(H1N1)pdm09 entstanden die höchsten direkten Kosten im Vergleich zu den anderen Altersgruppen. N2 - Influenza-associated respiratory disorders cause several hospitalizations and deaths every year. The pandemic virus Influenza A(H1N1)pdm09 caused several partly severe complications, especially for younger adults. In this study, we examined influenza-associated hospitalizations (IAH) regarding the course of disease in different age groups and to different virus types and -subtypes. Furthermore, we did a cost-analysis of influenza-associated hospitalizations. Especially severe influenza course occurred to adults aged 18-60 years with an underlying chronic condition, infected by influenza A(H1N1)pdm09. Also, pregnant women with IAH with influenza A(H1N1)pdm09 showed rare but severe complications. For patients aged 18-60 years infected by influenza A(H1N1)pdm09 occurred high costs compared to other age groups. KW - Influenza KW - Influenza A(H1N1)pdm09 KW - Schwangere KW - Komplikationen KW - direkte Kosten Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-199907 ER - TY - JOUR A1 - Kühnemundt, Johanna A1 - Leifeld, Heidi A1 - Scherg, Florian A1 - Schmitt, Matthias A1 - Nelke, Lena C. A1 - Schmitt, Tina A1 - Bauer, Florentin A1 - Göttlich, Claudia A1 - Fuchs, Maximilian A1 - Kunz, Meik A1 - Peindl, Matthias A1 - Brähler, Caroline A1 - Kronenthaler, Corinna A1 - Wischhusen, Jörg A1 - Prelog, Martina A1 - Walles, Heike A1 - Dandekar, Thomas A1 - Dandekar, Gudrun A1 - Nietzer, Sarah L. T1 - Modular micro-physiological human tumor/tissue models based on decellularized tissue for improved preclinical testing JF - ALTEX N2 - High attrition-rates entailed by drug testing in 2D cell culture and animal models stress the need for improved modeling of human tumor tissues. In previous studies our 3D models on a decellularized tissue matrix have shown better predictivity and higher chemoresistance. A single porcine intestine yields material for 150 3D models of breast, lung, colorectal cancer (CRC) or leukemia. The uniquely preserved structure of the basement membrane enables physiological anchorage of endothelial cells and epithelial-derived carcinoma cells. The matrix provides different niches for cell growth: on top as monolayer, in crypts as aggregates and within deeper layers. Dynamic culture in bioreactors enhances cell growth. Comparing gene expression between 2D and 3D cultures, we observed changes related to proliferation, apoptosis and stemness. For drug target predictions, we utilize tumor-specific sequencing data in our in silico model finding an additive effect of metformin and gefitinib treatment for lung cancer in silico, validated in vitro. To analyze mode-of-action, immune therapies such as trispecific T-cell engagers in leukemia, as well as toxicity on non-cancer cells, the model can be modularly enriched with human endothelial cells (hECs), immune cells and fibroblasts. Upon addition of hECs, transmigration of immune cells through the endothelial barrier can be investigated. In an allogenic CRC model we observe a lower basic apoptosis rate after applying PBMCs in 3D compared to 2D, which offers new options to mirror antigen-specific immunotherapies in vitro. In conclusion, we present modular human 3D tumor models with tissue-like features for preclinical testing to reduce animal experiments. KW - modular tumor tissue models KW - invasiveness KW - bioreactor culture KW - combinatorial drug predictions KW - immunotherapies Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-231465 VL - 38 ER - TY - THES A1 - Hilligardt [geb. Rück], Deborah T1 - Methylierung pro- und antiinflammatorischer T-Helfer-Zell-spezifischer Transkriptionsfaktoren bei ausgewählten Krankheitsbildern T1 - Methylation of pro- and anti-inflammatory t-helper cell specific transcription factors in different disease pattern N2 - Die Regulation krankheitsrelevanter Gene und deren Proteine über Veränderungen in der DNA-Methylierung stellen einen wichtigen und zugleich noch unzureichend erforschten Bereich bei Erkrankungen mit inflammatorischer Komponente dar. In dieser Arbeit wurde die Methylierung pro- und antiinflammatorischer Gene im hypoxischen Setting hervorgerufen durch Präeklampsie, Angsterkrankung und Inflammation bei Sklerodermie untersucht. Zur Bestimmung der prozentualen Methylierung wurde Pyrosequenzierung durchgeführt. Bei einem Teil der Proben erfolgte zusätzlich die Bestimmung der Genexpression mittels Real Time PCR. Bei Angsterkrankung zeigte sich eine signifikante Hypermethylierung am Promotor des Treg spezifischen Transkriptionsfaktors FOXP3. Daraus könnte eine beeinträchtigte Funktion der Tregs und somit eine erhöhte Komorbidität resultieren. In der Gruppe der an Sklerodermie erkrankten Personen zeigte sich entgegen den Erwartungen eine signifikant höhere RORC1 und RORC2 Methylierung. Eine Genexpressionsanalyse erbrachte eine signifikant niedrigere Expression von RORC bei Sklerodermie im Vergleich zu gesunden Kontrollen. Diese überraschenden Ergebnisse könnten der Methodik geschuldet sein. Auf eine Auftrennung der verschiedenen T-Zellen vor Messung der Methylierung wurde verzichtet. Plazentagewebe bei Präeklampsie zeigte eine signifikant geringere Methylierung am FOXP3 Promotor als Plazentagewebe von gesunden Schwangeren. Die Veränderbarkeit der DNA-Methylierung durch äußere Einflüsse und Medikamente stellt hierbei einen vielversprechenden Ansatzpunkt für zukünftige Therapien dar und sollte in weiteren Studien konkretisiert werden. N2 - Epigenetic research offers new insights about the regulation of gene activities and provides important knowledge of the pathogenesis of diseases. Hypoxic conditions through preeclampsia, panic disease and inflammation in systemic sclerosis were used to measure the methylation level of pro- and anti-inflammatory genes. The analyses were performed with PBMCs and placental tissue. To determine the methylation level pyrosequencing was used. Gene expression through real time PCR was additionally analyzed with part of the samples. A significant hypermethylation of the promoter of the Treg specific transcription factor FOXP3 in patients with panic diseases was shown. This could be a reason for the impaired function of Tregs in panic disorder and could cause the comorbidity of several diseases. Against expectations the transcription factor RORC was significantly higher methylated in patients with scleroderma and the gene expression was lower compared to the healthy control group. This surprising result might be caused through the used methods: t-cells were not divided into their subgroups. It could be possible that Th1- and Th2-cells are responsible for the hypermethylation of RORC. Placental tissue of patients with preeclampsia showed significant lower methylation levels of the FOXP3 promoter than tissue of healthy pregnant women. The convertibility of DNA methylation with external factors and pharmaceuticals is a promising approach for therapies and should be substantiated in future studies. KW - Methylierung KW - Epigenetik KW - Präeklampsie KW - Angststörung KW - Sklerodermie Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-249499 ER - TY - JOUR A1 - Scheer, Monika A1 - Vokuhl, Christian A1 - Blank, Bernd A1 - Hallmen, Erika A1 - von Kalle, Thekla A1 - Münter, Marc A1 - Wessalowski, Rüdiger A1 - Hartwig, Maite A1 - Sparber-Sauer, Monika A1 - Schlegel, Paul-Gerhardt A1 - Kramm, Christof M. A1 - Kontny, Udo A1 - Spriewald, Bernd A1 - Kegel, Thomas A1 - Bauer, Sebastian A1 - Kazanowska, Bernarda A1 - Niggli, Felix A1 - Ladenstein, Ruth A1 - Ljungman, Gustaf A1 - Jahnukainen, Kirsi A1 - Fuchs, Jörg A1 - Bielack, Stefan S. A1 - Klingebiel, Thomas A1 - Koscielniak, Ewa T1 - Desmoplastic small round cell tumors: Multimodality treatment and new risk factors JF - Cancer Medicine N2 - Background To evaluate optimal therapy and potential risk factors. Methods Data of DSRCT patients <40 years treated in prospective CWS trials 1997-2015 were analyzed. Results Median age of 60 patients was 14.5 years. Male:female ratio was 4:1. Tumors were abdominal/retroperitoneal in 56/60 (93%). 6/60 (10%) presented with a localized mass, 16/60 (27%) regionally disseminated nodes, and 38/60 (63%) with extraperitoneal metastases. At diagnosis, 23/60 (38%) patients had effusions, 4/60 (7%) a thrombosis, and 37/54 (69%) elevated CRP. 40/60 (67%) patients underwent tumor resection, 21/60 (35%) macroscopically complete. 37/60 (62%) received chemotherapy according to CEVAIE (ifosfamide, vincristine, actinomycin D, carboplatin, epirubicin, etoposide), 15/60 (25%) VAIA (ifosfamide, vincristine, adriamycin, actinomycin D) and, 5/60 (8%) P6 (cyclophosphamide, doxorubicin, vincristine, ifosfamide, etoposide). Nine received high-dose chemotherapy, 6 received regional hyperthermia, and 20 received radiotherapy. Among 25 patients achieving complete remission, 18 (72%) received metronomic therapies. Three-year event-free (EFS) and overall survival (OS) were 11% (±8 confidence interval [CI] 95%) and 30% (±12 CI 95%), respectively, for all patients and 26.7% (±18.0 CI 95%) and 56.9% (±20.4 CI 95%) for 25 patients achieving remission. Extra-abdominal site, localized disease, no effusion or ascites only, absence of thrombosis, normal CRP, complete tumor resection, and chemotherapy with VAIA correlated with EFS in univariate analysis. In multivariate analysis, significant factors were no thrombosis and chemotherapy with VAIA. In patients achieving complete remission, metronomic therapy with cyclophosphamide/vinblastine correlated with prolonged time to relapse. Conclusion Pleural effusions, venous thrombosis, and CRP elevation were identified as potential risk factors. The VAIA scheme showed best outcome. Maintenance therapy should be investigated further. KW - C-reactive protein KW - desmoplastic small round cell tumor KW - maintenance therapy KW - soft tissue sarcoma KW - Trousseau's syndrome Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-228444 VL - 8 IS - 2 ER - TY - JOUR A1 - Silwedel, Christine A1 - Speer, Christian P. A1 - Haarmann, Axel A1 - Fehrholz, Markus A1 - Claus, Heike A1 - Buttmann, Mathias A1 - Glaser, Kirsten T1 - Novel insights into neuroinflammation: bacterial lipopolysaccharide, tumor necrosis factor α, and Ureaplasma species differentially modulate atypical chemokine receptor 3 responses in human brain microvascular endothelial cells JF - Journal of Neuroinflammation N2 - Background: Atypical chemokine receptor 3 (ACKR3, synonym CXCR7) is increasingly considered relevant in neuroinflammatory conditions, in which its upregulation contributes to compromised endothelial barrier function and may ultimately allow inflammatory brain injury. While an impact of ACKR3 has been recognized in several neurological autoimmune diseases, neuroinflammation may also result from infectious agents, including Ureaplasma species (spp.). Although commonly regarded as commensals of the adult urogenital tract, Ureaplasma spp. may cause invasive infections in immunocompromised adults as well as in neonates and appear to be relevant pathogens in neonatal meningitis. Nonetheless, clinical and in vitro data on Ureaplasma-induced inflammation are scarce. Methods: We established a cell culture model of Ureaplasma meningitis, aiming to analyze ACKR3 variances as a possible pathomechanism in Ureaplasma-associated neuroinflammation. Non-immortalized human brain microvascular endothelial cells (HBMEC) were exposed to bacterial lipopolysaccharide (LPS) or tumor necrosis factor-α (TNF-α), and native as well as LPS-primed HBMEC were cultured with Ureaplasma urealyticum serovar 8 (Uu8) and U. parvum serovar 3 (Up3). ACKR3 responses were assessed via qRT-PCR, RNA sequencing, flow cytometry, and immunocytochemistry. Results: LPS, TNF-α, and Ureaplasma spp. influenced ACKR3 expression in HBMEC. LPS and TNF-α significantly induced ACKR3 mRNA expression (p < 0.001, vs. control), whereas Ureaplasma spp. enhanced ACKR3 protein expression in HBMEC (p < 0.01, vs. broth control). Co-stimulation with LPS and either Ureaplasma isolate intensified ACKR3 responses (p < 0.05, vs. LPS). Furthermore, stimulation wielded a differential influence on the receptor’s ligands. Conclusions: We introduce an in vitro model of Ureaplasma meningitis. We are able to demonstrate a pro-inflammatory capacity of Ureaplasma spp. in native and, even more so, in LPS-primed HBMEC, underlining their clinical relevance particularly in a setting of co-infection. Furthermore, our data may indicate a novel role for ACKR3, with an impact not limited to auto-inflammatory diseases, but extending to infection-related neuroinflammation as well. AKCR3-induced blood-brain barrier breakdown might constitute a potential common pathomechanism. KW - atypical chemokine receptor 3 KW - human brain microvascular endothelial cells KW - meningitis KW - neuroinflammation KW - Ureaplasma species Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-175952 VL - 15 IS - 156 ER - TY - JOUR A1 - Fiore, Piera Filomena A1 - Vacca, Paola A1 - Tumino, Nicola A1 - Besi, Francesca A1 - Pelosi, Andrea A1 - Munari, Enrico A1 - Marconi, Marcella A1 - Caruana, Ignazio A1 - Pistoia, Vito A1 - Moretta, Lorenzo A1 - Azzarone, Bruno T1 - Wilms' tumor primary cells display potent immunoregulatory properties on NK cells and macrophages JF - Cancers N2 - The immune response plays a crucial defensive role in cancer growth and metastasis and is a promising target in different tumors. The role of the immune system in Wilm’s Tumor (WT), a common pediatric renal malignancy, is still to be explored. The characterization of the immune environment in WT could allow the identification of new therapeutic strategies for targeting possible inhibitory mechanisms and/or lowering toxicity of the current treatments. In this study, we stabilized four WT primary cultures expressing either a blastematous (CD56\(^+\)/CD133\(^−\)) or an epithelial (CD56\(^−\)/CD133\(^+\)) phenotype and investigated their interactions with innate immune cells, namely NK cells and monocytes. We show that cytokine-activated NK cells efficiently kill WT cells. However, after co-culture with WT primary cells, NK cells displayed an impaired cytotoxic activity, decreased production of IFNγ and expression of CD107a, DNAM-1 and NKp30. Analysis of the effects of the interaction between WT cells and monocytes revealed their polarization towards alternatively activated macrophages (M2) that, in turn, further impaired NK cell functions. In conclusion, we show that both WT blastematous and epithelial components may contribute directly and indirectly to a tumor immunosuppressive microenvironment that is likely to play a role in tumor progression. KW - Wilm's tumor KW - NK cells KW - macrophages KW - tumor microenvironment KW - Wilms' tumor Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-222981 SN - 2072-6694 VL - 13 IS - 2 ER - TY - JOUR A1 - Ruf, Katharina A1 - Beer, Meinrad A1 - Köstler, Herbert A1 - Weng, Andreas Max A1 - Neubauer, Henning A1 - Klein, Alexander A1 - Platek, Kathleen A1 - Roth, Kristina A1 - Beneke, Ralph A1 - Hebestreit, Helge T1 - Size-adjusted muscle power and muscle metabolism in patients with cystic fibrosis are equal to healthy controls – a case control study JF - BMC Pulmonary Medicine N2 - Background Skeletal muscle function dysfunction has been reported in patients with cystic fibrosis (CF). Studies so far showed inconclusive data whether reduced exercise capacity is related to intrinsic muscle dysfunction in CF. Methods Twenty patients with CF and 23 age-matched controls completed an incremental cardiopulmonary cycling test. Further, a Wingate anaerobic test to assess muscle power was performed. In addition, all participants completed an incremental knee-extension test with 31P magnetic resonance spectroscopy to assess muscle metabolism (inorganic phosphate (Pi) and phosphocreatinine (PCr) as well as intracellular pH). In the MRI, muscle cross-sectional area of the M. quadriceps (qCSA) was also measured. A subgroup of 15 participants (5 CF, 10 control) additionally completed a continuous high-intensity, high-frequency knee-extension exercise task during 31P magnetic resonance spectroscopy to assess muscle metabolism. Results Patients with CF showed a reduced exercise capacity in the incremental cardiopulmonary cycling test (VO2peak: CF 77.8 ± 16.2%predicted (36.5 ± 7.4 ml/qCSA/min), control 100.6 ± 18.8%predicted (49.1 ± 11.4 ml/qCSA/min); p < 0.001), and deficits in anaerobic capacity reflected by the Wingate test (peak power: CF 537 ± 180 W, control 727 ± 186 W; mean power: CF 378 ± 127 W, control 486 ± 126 W; power drop CF 12 ± 5 W, control 8 ± 4 W. all: p < 0.001). In the knee-extension task, patients with CF achieved a significantly lower workload (p < 0.05). However, in a linear model analysing maximal work load of the incremental knee-extension task and results of the Wingate test, respectively, only muscle size and height, but not disease status (CF or not) contributed to explaining variance. In line with this finding, no differences were found in muscle metabolism reflected by intracellular pH and the ratio of Pi/PCr at submaximal stages and peak exercise measured through MRI spectroscopy. Conclusions The lower absolute muscle power in patients with CF compared to controls is exclusively explained by the reduced muscle size in this study. No evidence was found for an intrinsic skeletal muscle dysfunction due to primary alterations of muscle metabolism. KW - Cystic fibrosis KW - Exercise capacity KW - MRI spectroscopy KW - Muscle power KW - Phosphorylation KW - Lung disease, KW - Muscle function Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-200981 VL - 19 ER - TY - JOUR A1 - Fehrholz, Markus A1 - Glaser, Kirsten A1 - Speer, Christian P. A1 - Seidenspinner, Silvia A1 - Ottensmeier, Barbara A1 - Kunzmann, Steffen T1 - Caffeine modulates glucocorticoid-induced expression of CTGF in lung epithelial cells and fibroblasts JF - Respiratory Research N2 - Background: Although caffeine and glucocorticoids are frequently used to treat chronic lung disease in preterm neonates, potential interactions are largely unknown. While anti-inflammatory effects of glucocorticoids are well defined, their impact on airway remodeling is less characterized. Caffeine has been ascribed to positive effects on airway inflammation as well as remodeling. Connective tissue growth factor (CTGF, CCN2) plays a key role in airway remodeling and has been implicated in the pathogenesis of chronic lung diseases such as bronchopulmonary dysplasia (BPD) in preterm infants. The current study addressed the impact of glucocorticoids on the regulation of CTGF in the presence of caffeine using human lung epithelial and fibroblast cells. Methods: The human airway epithelial cell line H441 and the fetal lung fibroblast strain IMR-90 were exposed to different glucocorticoids (dexamethasone, budesonide, betamethasone, prednisolone, hydrocortisone) and caffeine. mRNA and protein expression of CTGF, TGF-β1-3, and TNF-α were determined by means of quantitative real-time PCR and immunoblotting. H441 cells were additionally treated with cAMP, the adenylyl cyclase activator forskolin, and the selective phosphodiesterase (PDE)-4 inhibitor cilomilast to mimic caffeine-mediated PDE inhibition. Results: Treatment with different glucocorticoids (1 μM) significantly increased CTGF mRNA levels in H441 (p < 0.0001) and IMR-90 cells (p < 0.01). Upon simultaneous exposure to caffeine (10 mM), both glucocorticoid-induced mRNA and protein expression were significantly reduced in IMR-90 cells (p < 0.0001). Of note, 24 h exposure to caffeine alone significantly suppressed basal expression of CTGF mRNA and protein in IMR-90 cells. Caffeine-induced reduction of CTGF mRNA expression seemed to be independent of cAMP levels, adenylyl cyclase activation, or PDE-4 inhibition. While dexamethasone or caffeine treatment did not affect TGF-β1 mRNA in H441 cells, increased expression of TGF-β2 and TGF-β3 mRNA was detected upon exposure to dexamethasone or dexamethasone and caffeine, respectively. Moreover, caffeine increased TNF-α mRNA in H441 cells (6.5 ± 2.2-fold, p < 0.05) which has been described as potent inhibitor of CTGF expression. Conclusions: In addition to well-known anti-inflammatory features, glucocorticoids may have adverse effects on long-term remodeling by TGF-β1-independent induction of CTGF in lung cells. Simultaneous treatment with caffeine may attenuate glucocorticoid-induced expression of CTGF, thereby promoting restoration of lung homeostasis. KW - airway remodeling KW - fibrosis KW - bronchopulmonary dysplasia KW - caffeine KW - CCN2 KW - CTGF KW - glucocorticoids KW - H441 KW - IMR-90 Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-157672 VL - 18 IS - 51 ER - TY - JOUR A1 - Nothhaft, Matthias A1 - Klepper, Joerg A1 - Kneitz, Hermann A1 - Meyer, Thomas A1 - Hamm, Henning A1 - Morbach, Henner T1 - Hemorrhagic bullous Henoch-Schönlein Purpura: case report and review of the literature JF - Frontiers in Pediatrics N2 - Henoch-Schönlein Purpura (HSP) or IgA vasculitis is the most common systemic vasculitis of childhood and may affect skin, joints, gastrointestinal tract, and kidneys. Skin manifestations of HSP are characteristic and include a non-thrombocytopenic palpable purpura of the lower extremities and buttocks. Rarely, HSP may initially present as or evolve into hemorrhagic vesicles and bullae. We present an otherwise healthy 5-year-old boy with an acute papulovesicular rash of both legs and intermittent abdominal pain. After a few days the skin lesions rapidly evolved into palpable purpura and hemorrhagic bullous lesions of variable size and severe hemorrhagic HSP was suspected. A histological examination of a skin biopsy showed signs of a small vessel leukocytoclastic vasculitis limited to the upper dermis and direct immunofluorescence analysis revealed IgA deposits in vessel walls, compatible with HSP. To further characterize the clinical picture and treatment options of bullous HSP we performed an extensive literature research and identified 41 additional pediatric patients with bullous HSP. Two thirds of the reported patients were treated with systemic corticosteroids, however, up to 25% of the reported patients developed skin sequelae such as hyperpigmentation and/or scarring. The early use of systemic corticosteroids has been discussed controversially and suggested in some case series to be beneficial by reducing the extent of lesions and minimizing sequelae of disease. Our patient was treated with systemic corticosteroids tapered over 5 weeks. Fading of inflammation resulted in healing of most erosions, however, a deep necrosis developing from a large blister at the dorsum of the right foot persisted so that autologous skin transplantation was performed. Re-examination 11 months after disease onset showed complete clinical remission with re-epithelialization but also scarring of some affected areas. KW - henoch-schönlein purpura KW - vasculitis KW - hemorrhagic KW - bullae KW - children Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-201435 VL - 6 ER - TY - THES A1 - Krakow, Sören T1 - CD14-Reexpression definiert einen immunregulatorischen Phänotyp Monozyten-gereifter Zellen nach IL-10/R848-Stimulation T1 - Re-expression of CD14 in Response to a Combined IL-10/TLR Stimulus Defines Monocyte-Derived Cells With an Immunoregulatory Phenotype N2 - Dendritische Zellen können als antigenpräsentierende Zellen sowohl immunogene als auch tolerogene Funktionen im Immunsystem wahrnehmen und werden in der Therapie von Tumorerkrankungen und Autoimmunerkrankungen eingesetzt. IL-10 gilt als Induktor tolerogener dendritischer Zellen. Diese werden in der Literatur oft als unreif bezeichnet und stehen im Gegensatz zu den reifen immunogenen dendritischen Zellen, die durch die Expression des Reifungsmarkers CD83 gekennzeichnet sind. Ausdifferenzierte dendritische Zellen exprimieren zudem das Antigen CD86, das der T-Zell-Aktivierung dient. In der vorliegenden Arbeit wurde der Einfluss von IL-10 auf den Reifungsprozess dendritischer Zellen in vitro untersucht. Zur Generierung unreifer dendritischer Zellen wurden humane Monozyten nach etabliertem Protokoll mit IL-4 und GM-CSF stimuliert. Nach anschließender IL-10-Stimulation, insbesondere in Kombination mit einem TLR-Agonisten, bildeten sich zwei exklusive Zellpopulationen: eine CD14+ Population und eine CD83+ Population. Unreife CD14-CD83- dendritische Zellen reexprimierten einerseits CD14 oder exprimierten andererseits CD83. Dabei zeigte sich, dass das kostimulierende Antigen CD86 gleichermaßen sowohl mit als auch ohne IL-10-Inkubation hoch exprimiert wurde und IL-10 folglich keinen zusätzlichen Einfluss auf dessen Expression hat. Insgesamt waren Veränderungen bezüglich der Oberflächenantigene, die bei der Betrachtung der Gesamtheit aller Zellen auffallen, auf eine quantitative Verschiebung der beiden Zellpopulationen zurückzuführen. IL-10 beeinflusst also nicht direkt einzelne kostimulierende oder inhibitorische Moleküle, sondern beeinflusst den Anteil der CD14+ Zellen gegenüber den CD83+ dendritischen Zellen. Funktionell betrachtet zeigten die CD14+ Zellen eine gesteigerte Makropinozytose im Gegensatz zu den reifen CD83+ dendritischen Zellen. Zusammenfassend führt IL-10 zu einer Reexpression von CD14 auf unreifen dendritischen Zellen und aktiviert einen alternativen Differenzierungsweg. Die CD14+ Zellen weisen einen stabilen immunregulatorischen Phänotyp auf und unterscheiden sich somit von reifen dendritischen Zellen, die nach Inkubation mit IL-10 nicht reguliert werden. Damit muss die Begrifflichkeit und Klassifikation tolerogener dendritischer Zellen weiter diskutiert werden. N2 - Dendritic cells (DCs) are antigen-presenting cells and play a critical role in innate and acquired immunity as far as they can initiate and boost T cell responses. They also maintain the balance between tolerance and immunity. However, the phenotypic and functional diversity of dendritic cell subsets still are the main subject of research. Interleukin 10 is a central regulator of the antigen-presenting function of myeloid cells. It exerts immunomodulatory effects in vivo and induces a regulatory phenotype in monocyte-derived cells in vitro. In this study, the phenotype and function of monocytic cells are analyzed in vitro in relation to the cytokine milieu and the timing of TLR-based activation. In GM-CSF/IL-4 cultured human monocytic cells, two mutually exclusive cell populations - arising from undifferentiated cells - are identified: CD83+ fully activated dendritic cells and CD14+ macrophage like cells. Re-expression of CD14 occurs primarily after a sequential trigger with a TLR signal following IL-10 preincubation. This cell population with re-expressed CD14 substantially differs in phenotype and function from the CD83+ cells. Detailed analysis of individual subpopulations reveals that exogenous IL-10 is critical for inducing the shift toward the CD14+ population but does not affect individual changes in marker expression or cell function in most cases. Thus, plasticity of CD14 expression, defining a subset of immunoregulatory cells, is highly relevant for the composition of cellular products (such as DC vaccines) as it affects the function of the total product. KW - Dendritische Zelle KW - Monozyt KW - Makrophage KW - IL-10 Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-224280 ER -