TY - THES A1 - Steigerwald, Mario T1 - Die Rolle von dendritischen Zellen und Chemokinen bei der Regulation der Immunantwort gegen Erreger der kutanen Leishmaniose T1 - The role of dendritic cells and chemokines in the regulation of the immune response against pathogens of cutaneous leishmaniasis N2 - Dendritische Zellen stellen ein essentielles Bindeglied zwischen angeborener und adaptiver Immunität dar. Sie stammen aus dem Knochenmark und bilden ein Netzwerk von heterogenen Zellpopulationen. In peripheren Geweben liegen sie als unreife Zellen mit einem hohen Potential zur Aufnahme und Prozessierung von Mikroorganismen vor. Nach der Aufnahme von eindringenden Mikroorganismen beginnen dendritische Zellen jedoch, sich von einem prozessierenden in ein präsentierendes Stadium zu differenzieren, und wandern zu den sekundären lymphatischen Organen, um dort den naïven T-Zellen die mikrobiellen Antigene zu präsentieren. Die gerichtete Wanderung von dendritischen Zellen ist hierbei ein zentraler Bestandteil der immunstimulatorischen und -modulatorischen Funktion dieser Zellen. Eine essentielle Rolle bei diesem Migrationsverhalten spielen chemotaktische Zytokine (Chemokine). Chemokine sind Proteine mit einem niedermolekularen Gewicht (8-10 kDa), welche anhand ihres strukturellen Cysteinmotifs in vier Gruppen unterteilt und entweder als CXC-, CC-, C- und CX3C- Chemokine oder als α-, β-, δ- und γ-Chemokine bezeichnet werden. Arbeiten der eigenen Arbeitsgruppe am Modell der kutanen Leishmaniose haben jedoch gezeigt, dass die Funktion von Chemokinen weitaus mehr umfasst, als lediglich die zielgerichtete Steuerung der Migration immunologisch wichtiger Zellen. So konnte in diesen Studien nicht nur gezeigt werden, dass das Muster der Expression von Chemokinen mit der Schwere des Krankheitsbildes korreliert, sondern auch, dass das Chemokin CCL2/MCP-1 in der Lage ist, einen direkten Einfluss auf die intrazelluläre Erregerabwehr zu nehmen. Diese Arbeiten bezogen sich jedoch auf humane Hautbiopsien und aus Humanblut isolierten Zellen. Eine detaillierte Analyse des Infektionsverlaufes unter definierten Bedingungen (konstante Infektionsdosis und -art, kontrollierte Infektionsdauer, Verwendung von klonierten Parasiten, einheitlicher genetischer Hintergrund des Wirts) ist bei Patienten jedoch nicht möglich. Deshalb wurde die experimentelle Leishmanieninfektion von Inzuchtmäusen (suszeptible BALB/c- und resistente C57BL/6-Mäuse) herangezogen, um die Kinetik der Chemokinexpression und deren Korrelation mit dem Verlauf der kutanen Leishmaniose zu bestimmen. Die Arbeiten dieser Doktorarbeit zeigen erstmals, dass ebenso wie im humanen System bei der experimentellen Leishmaniose in der Maus die Fähigkeit zur Abwehr dieses Erregers mit einer verstärkten Expression von CCL2/MCP-1 in der infizierten Haut korreliert. Des Weiteren konnte zwar eine CCL2/MCP-1-induzierte leishmanizide Wirkung in murinen Makrophagen festgestellt werden, ein vergleichbarer Effekt blieb bei Langerhans-Zellen, den dendritischen Zellen der Haut, jedoch aus. Weiterhin sollte der Einfluss einer Leishmanieninfektion auf das CCL2/MCP-1- induzierte Wanderungsverhalten von Langerhans-Zellen untersucht werden, da aus der Literatur bekannt ist, dass das Chemokin CCL2/MCP-1 die Migration dendritischer Zellen induziert. Die Studien der Doktorarbeit ergaben hierbei, dass eine Infektion mit Leishmania major zu einer signifikanten Verminderung der durch CL2/MCP-1 oder CCL3/MIP-1α induzierten Migration von Langerhans-Zellen führt. Eine mögliche Ursache für dieses Resultat war hierbei in dem Einfluss einer Infektion mit Leishmanien auf die Expression von Chemokinrezeptoren in dendritischen Zellen zu finden. Anschließende Untersuchungen der mRNA-Expression dieser Rezeptoren konnten diese Vermutung bestätigen. So wurde nach einer Infektion von dendritischen Zellen mit L. major eine Reduktion der mRNA-Expression der Chemokinrezeptoren CCR2 und CCR5 festgestellt. Anschließende FACS-Analysen und Studien mit Hilfe des konfokalen Lasermikroskops führten zu einem ähnlichen Resultat. Interessanterweise bewirkt die Infektion mit L. major andererseits eine Hochregulation der mRNA-Expression von CCR7 in dendritischen Zellen. Von diesem Rezeptor ist bekannt, dass er die Chemokine CCL19/MIP-3β und CCL21/6Ckine, welche im Lymphknoten konstitutiv exprimiert werden, mit großer Affinität bindet und für die zielgerichtete Migration dendritischer Zellen in dieses Organ essentiell ist. Weitere Versuche ergaben zudem eine verstärkte CXCL10/IP-10-mRNA-Expression in dendritischen Zellen aus L. major-resistenten C57BL/6-Mäusen, welche bei dendritischen Zellen aus BALB/c-Mäusen nicht festgestellt worden ist. Zusammenfassend konnte in dieser Doktorarbeit gezeigt werden, dass eine Infektion dendritischer Zellen mit L. major sowohl bei suszeptiblen als auch bei resistenten Mäusen zu einer Modulation der Expression von Chemokinrezeptoren führt, die sowohl für die Lokalisation der Zellen in den entzündeten Hautarealen verantwortlich sind (CCR2 und CCR5), als auch ihre Wanderung in die Lymphknoten steuern CCR7). Darüber hinaus lässt die wirtsspezifische Modulation des Chemokins CXCL10/IP-10 in resistenten Tieren vermuten, dass sie zur Kontrolle der Infektion beiträgt. N2 - Dendritic cells (DC) represent an essential link between innate and adaptive immunity. DC are bone marrow-derived and form a network of heterogeneous cell populations. In peripheral tissue they are in an immature state, with a high potential for taking up and processing microorganisms. After taking up invading pathogens, DC differentiate from a “processing” into a “presenting” stage, while migrating to the secondary lymphoid organs in order to present microbial antigens to naïve T cells. The directed migration of dendritic cells is a central component in the immunostimulatory and modulatory function of these cells. Chemotactic cytokines (chemokines) are critical for these migratory pathways. Chemokines are proteins of low molecular weight (8-10 kDa) and can be classified into four groups on the basis of a cysteine structural motif, known as either the CXC, CC, C and CX3C or the α, β, δ and γ subfamilies. Studies using the model of cutaneous leishmaniasis have shown, however, that the function of chemokines is much wider than only to control the migratory pathway of immunologically relevant cells. These previous studies demonstrated not only the correlation between the chemokine expression pattern and the course of the infection, but also the direct influence of the chemokine CCL2/MCP-1 on the intracellular defence of pathogens. However, these studies were based on skin lesions from patients and human monocytes. A more detailed analysis of the infection process under more defined conditions (constant infection dose, defined duration of infection, use of cloned parasites, uniform genetic background of the host) is not possible with human material. Therefore, the experimental Leishmania model with inbred mice (susceptible BALB/c and resistant C57BL/6 mice) was used to determine the kinetics of the chemokine expression and their correlation with the course of cutaneous leishmaniasis. The studies of this work demonstrated for the first time that the expression of CCL2/MCP-1 during experimental infection with Leishmania major in mice correlates with the capability to control this pathogen. Moreover, a CCL2/MCP-1 induced leishmanicidal effect on murine macrophages could be determined, but a comparable effect was not observed with Langerhans cells, the dendritic cells of the skin. Furthermore, in this work the influence of an infection with Leishmania on the CCL2/MCP-1-induced migration of Langerhans cells were examined, since it is known from the literature that CCL2/MCP-1 induces dendritic cell migration. The studies of this work demonstrated that an infection of Langerhans cells with L. major leads to a significant reduction of the CCL2/MCP-1- or CCL3/MIP-1α-induced migration rate of these cells. A possible reason for this result may be the influence of an infection with Leishmania on the chemokine receptor expression by dendritic cells. Further investigations of the mRNA expression of these receptors could confirm this assumption. They demonstrated reductions in the mRNA expression of the chemokine receptors CCR2 and CCR5 after an infection of dendritic cells with L. major. Moreover, FACS analysis and studies using confocal laser microscopy led to similar results and showed that the L. major-induced alterations in mRNA expression correlate with those at the protein level. Interestingly, a strong upregulation of CCR7 mRNA expression could be detected after infection of dendritic cells. This receptor is well known for binding the chemokines CCL19/MIP-3β and CCL21/6Ckine, which both are constitutively expressed on the lymph node, with high affinity. Moreover, this receptor is essential for a directed movement of dendritic cells to that organ. Further studies also showed an upregulation of the CXCL10/IP-10-mRNA expression in dendritic cells from L. major-resistant C57BL/6 mice, whereas it could not be detected in dendritic cells from susceptible BALB/c mice. In summary, this work demonstrated that an L. major-infection of dendritic cells from resistant mice as well as susceptible mice leads to a modulation in the expression of chemokine receptors that are responsible for the localisation of those cells into the inflammatory tissue (CCR2 and CCR5) and the migration of dendritic cells to the lymph node (CCR7). Moreover, the host-specific modulation of the chemokine CXCL10/IP-10 in resistant animals seems to play a role in the control of the infection. KW - Leishmaniose KW - Dendritische Zelle KW - Chemokine KW - Immunreaktion KW - dendritische Zelle KW - kutane Leishmaniose KW - Chemokine KW - Immunantwort KW - dendritic cells KW - cutane leishmaniasis KW - chemokine KW - immune response Y1 - 2005 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-13852 ER - TY - THES A1 - Bittner, Thomas T1 - Immunhistologische Charakterisierung immunkompetenter Zellen in Plattenepithelkarzinomen des oberen Aerodigestivtraktes T1 - Immunohistochemical characterisation of immunocompetent cells in squamous cell carcinomas of the upper aerodigestive tract N2 - Ziel der vorliegenden Studie war es, die Beziehung zwischen Expression der Basalmembran und der damit einhergehen Veränderung des Immunzelleninfiltrates zu untersuchen. Eine Reihe von 23 Plattenepithelkarzinomen des Larynx und Hypopharynx wurden lichtmikroskopisch untersucht. Hierzu wurde eine immunhistochemischen Färbung verwendet mit monoklonalen Antikörpern gegen folgende Membranantigene: CD 1a, CD 4, CD 8, Pan B, CD 14, ICAM 1, LFA, HLA-DR, Kollagen 4. Die immunhistochemische Analyse wurde bezogen auf den Differenzierungsgrad des Tumors, definiert durch die Ausprägung der Basalmembran. Epithel und Stroma wurden getrennt ausgezählt. Der Verlust der Basalmembran wurde von einer Veränderung in der Zusammensetzung des Immunzelleninfiltrates begleitet: HLA-DR und LFA nahm in beiden Kompartimenten ab. Pan B und CD 14 zeigten eine Parallelität im Färbeverhalten. Pan B zeigte die stärkste Färbung in Plattenepithelkarzinomen mit annähernd ununterbrochener Basalmembran. CD 8 zeigte die stärksteFärbung in Plattenepithelkarzinomen mit lückenhafter, aber noch vorhandener Basalmembran. Die CD 8 positiven Zellen zeigten stellenweise engen Kontakt zur Basalmembran, stellenweise keimzentrumsähnliche Anordnungen. Wir schlussfolgern, dass bei gut differenzierten Tumoren das B-Zell System, bei mäßig differenzierten das T-Zell System eine gewisse Rolle spielt. Der Übergangszone Tumor-Stroma scheint eine besondere immunologische Bedeutung zu zukommen. N2 - The objective of the present study was to evaluate the relationship between the expression of the basement Membrane (BM) in SCC and the relative proportions of selective immune cell populations. A series of 23 SCC of the Larynx and the Hypopharynx was assessed by light microscopy and using immunohistochemical staining with monoclonal antibodies directed against the following cell surface antigens: CD 1a, CD 4, CD 8, Pan B, CD 14, ICAM 1, LFA, HLA-DR, Collagen 4. Immunhistochemical analysis of the tumor infiltrating cells was performed and correlated with the transformation level as defined by the grade of expression of BM. The loss of BM was accompanied by a change in immune infiltrate: HLA-DR and LFA decreased in both compartiments. The B cells stained in parallel with CD 14 positive cells. Pan B showed the highest levels in SCC with a nearly continous BM. CD 8 peaked in tumors with a leaky but still present BM. The CD 8 cells showed sometimes tight junction to the BM, sometimes formations similar to germinal centers. We conclude that in well differentiated SCC the B cell system plays a certain role, in middle differentiated the T cell system. The epithelio-stromal junction zone seems to play an important immunologic role. KW - Plattenepithelkarzinom KW - immunkompetente Zellen KW - monoklonale Antikörper KW - Immunantwort KW - squamous cell carcinoma KW - immunocompetent cells KW - monoclonal antibodies KW - immune response Y1 - 2005 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-12087 ER - TY - JOUR A1 - Azzami, Klara A1 - Ritter, Wolfgang A1 - Tautz, Jürgen A1 - Beier, Hildburg T1 - Infection of honey bees with acute bee paralysis virus does not trigger humoral or cellular immune responses JF - Archives of Virology N2 - We have studied the responses of honey bees at different life stages (Apis mellifera) to controlled infection with acute bee paralysis virus and have identified the haemolymph of infected larvae and adult worker bees as the compartment where massive propagation of ABPV occurs. Insects respond with a broad spectrum of induced innate immune reactions to bacterial infections, whereas defence mechanisms based on RNA interference play a major role in antiviral immunity. In this study, we have determined that honey bee larvae and adult workers do not produce a humoral immune reaction upon artificial infection with ABPV, in contrast to control individuals challenged with Escherichia coli. ABPV-infected bees produced neither elevated levels of specific antimicrobial peptides (AMPs), such as hymenoptaecin and defensin, nor any general antimicrobial activity, as revealed by inhibition-zone assays. Additionally, adult bees did not generate melanised nodules upon ABPV infection, an important cellular immune function activated by bacteria and viruses in some insects. Challenge of bees with both ABPV and E. coli showed that innate humoral and cellular immune reactions are induced in mixed infections, albeit at a reduced level. KW - chemosensory protein KW - bee larva KW - adult bee KW - honey bee KW - larva KW - work bee KW - infected bee KW - immune response KW - young work bee KW - capsid protein KW - abdominal KW - tergite KW - haemolymph KW - sample KW - Imd pathway KW - worker bee larva KW - antimicrobial KW - peptide Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-126863 VL - 157 IS - 4 ER - TY - JOUR A1 - Schwarz, Tobias A1 - Remer, Katharina A. A1 - Nahrendorf, Wiebke A1 - Masic, Anita A1 - Siewe, Lisa A1 - Müller, Werner A1 - Roers, Axel A1 - Moll, Heidrun T1 - T Cell-Derived IL-10 Determines Leishmaniasis Disease Outcome and Is Suppressed by a Dendritic Cell Based Vaccine JF - PLoS Pathogens N2 - Abstract In the murine model of Leishmania major infection, resistance or susceptibility to the parasite has been associated with the development of a Th1 or Th2 type of immune response. Recently, however, the immunosuppressive effects of IL-10 have been ascribed a crucial role in the development of the different clinical correlates of Leishmania infection in humans. Since T cells and professional APC are important cellular sources of IL-10, we compared leishmaniasis disease progression in T cell-specific, macrophage/neutrophil-specific and complete IL-10-deficient C57BL/6 as well as T cell-specific and complete IL-10-deficient BALB/c mice. As early as two weeks after infection of these mice with L. major, T cell-specific and complete IL-10-deficient animals showed significantly increased lesion development accompanied by a markedly elevated secretion of IFN-γ or IFN-γ and IL-4 in the lymph nodes draining the lesions of the C57BL/6 or BALB/c mutants, respectively. In contrast, macrophage/neutrophil-specific IL-10-deficient C57BL/6 mice did not show any altered phenotype. During the further course of disease, the T cell-specific as well as the complete IL-10-deficient BALB/c mice were able to control the infection. Furthermore, a dendritic cell-based vaccination against leishmaniasis efficiently suppresses the early secretion of IL-10, thus contributing to the control of parasite spread. Taken together, IL-10 secretion by T cells has an influence on immune activation early after infection and is sufficient to render BALB/c mice susceptible to an uncontrolled Leishmania major infection. Author Summary The clinical symptoms caused by infections with Leishmania parasites range from self-healing cutaneous to uncontrolled visceral disease and depend not only on the parasite species but also on the type of the host's immune response. It is estimated that 350 million people worldwide are at risk, with a global incidence of 1–1.5 million cases of cutaneous and 500,000 cases of visceral leishmaniasis. Murine leishmaniasis is the best-characterized model to elucidate the mechanisms underlying resistance or susceptibility to Leishmania major parasites in vivo. Using T cell-specific and macrophage-specific mutant mice, we demonstrate that abrogating the secretion of the immunosuppressive cytokine IL-10 by T cells is sufficient to render otherwise susceptible mice resistant to an infection with the pathogen. The healing phenotype is accompanied by an elevated specific inflammatory immune response very early after infection. We further show that dendritic cell-based vaccination against leishmaniasis suppresses the early secretion of IL-10 following challenge infection. Thus, our study unravels a molecular mechanism critical for host immune defense, aiding in the development of an effective vaccine against leishmaniasis. KW - cytokines KW - mouse models KW - T cells KW - lymph nodes KW - leishmania major KW - secretion KW - parasitic diseases KW - immune response Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-130385 VL - 9 IS - 6 ER - TY - JOUR A1 - Brehm, Klaus A1 - Komguep Nono, Justin A1 - Lutz, Manfred B. T1 - EmTIP, a T-Cell Immunomodulatory Protein Secreted by the Tapeworm Echinococcus multilocularis Is Important N2 - Background Alveolar echinococcosis (AE), caused by the metacestode of the tapeworm Echinococcus multilocularis, is a lethal zoonosis associated with host immunomodulation. T helper cells are instrumental to control the disease in the host. Whereas Th1 cells can restrict parasite proliferation, Th2 immune responses are associated with parasite proliferation. Although the early phase of host colonization by E. multilocularis is dominated by a potentially parasitocidal Th1 immune response, the molecular basis of this response is unknown. Principal Findings We describe EmTIP, an E. multilocularis homologue of the human T-cell immunomodulatory protein, TIP. By immunohistochemistry we show EmTIP localization to the intercellular space within parasite larvae. Immunoprecipitation and Western blot experiments revealed the presence of EmTIP in the excretory/secretory (E/S) products of parasite primary cell cultures, representing the early developing metacestode, but not in those of mature metacestode vesicles. Using an in vitro T-cell stimulation assay, we found that primary cell E/S products promoted interferon (IFN)-γ release by murine CD4+ T-cells, whereas metacestode E/S products did not. IFN-γ release by T-cells exposed to parasite products was abrogated by an anti-EmTIP antibody. When recombinantly expressed, EmTIP promoted IFN-γ release by CD4+ T-cells in vitro. After incubation with anti-EmTIP antibody, primary cells showed an impaired ability to proliferate and to form metacestode vesicles in vitro. Conclusions We provide for the first time a possible explanation for the early Th1 response observed during E. multilocularis infections. Our data indicate that parasite primary cells release a T-cell immunomodulatory protein, EmTIP, capable of promoting IFN-γ release by CD4+ T-cells, which is probably driving or supporting the onset of the early Th1 response during AE. The impairment of primary cell proliferation and the inhibition of metacestode vesicle formation by anti-EmTIP antibodies suggest that this factor fulfills an important role in early E. multilocularis development within the intermediate host. KW - T Cells KW - primary cells KW - parasitic diseases KW - vesicles KW - larvae KW - immunoprecipitation KW - host-pathogen interactions KW - immune response Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-111407 ER - TY - JOUR A1 - Arndt, Andreas A1 - Hoffacker, Peter A1 - Zellmer, Konstantin A1 - Goecer, Oktay A1 - Recks, Mascha S. A1 - Kuerten, Stefanie T1 - Conventional Housing Conditions Attenuate the Development of Experimental Autoimmune Encephalomyelitis JF - PLoS ONE N2 - BACKGROUND: The etiology of multiple sclerosis (MS) has remained unclear, but a causative contribution of factors outside the central nervous system (CNS) is conceivable. It was recently suggested that gut bacteria trigger the activation of CNS-reactive T cells and the development of demyelinative disease. METHODS: C57BL/6 (B6) mice were kept either under specific pathogen free or conventional housing conditions, immunized with the myelin basic protein (MBP)-proteolipid protein (PLP) fusion protein MP4 and the development of EAE was clinically monitored. The germinal center size of the Peyer's patches was determined by immunohistochemistry in addition to the level of total IgG secretion which was assessed by ELISPOT. ELISPOT assays were also used to measure MP4-specific T cell and B cell responses in the Peyer's patches and the spleen. Ear swelling assays were performed to determine the extent of delayed-type hypersensitivity reactions in specific pathogen free and conventionally housed mice. RESULTS: In B6 mice that were actively immunized with MP4 and kept under conventional housing conditions clinical disease was significantly attenuated compared to specific pathogen free mice. Conventionally housed mice displayed increased levels of IgG secretion in the Peyer's patches, while the germinal center formation in the gut and the MP4-specific TH17 response in the spleen were diminished after immunization. Accordingly, these mice displayed an attenuated delayed type hypersensitivity (DTH) reaction in ear swelling assays. CONCLUSIONS: The data corroborate the notion that housing conditions play a substantial role in the induction of murine EAE and suggest that the presence of gut bacteria might be associated with a decreased immune response to antigens of lower affinity. This concept could be of importance for MS and calls for caution when considering the therapeutic approach to treat patients with antibiotics." KW - B cells KW - secretion KW - multiple sclerosis KW - enzyme-linked immunoassays KW - Peyer's patches KW - gut bacteria KW - T cells KW - immune response Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-119603 VL - 9 IS - 6 ER - TY - JOUR A1 - Wong, David A1 - Winter, Oliver A1 - Hartig, Christina A1 - Siebels, Svenja A1 - Szyska, Martin A1 - Tiburzy, Benjamin A1 - Meng, Lingzhang A1 - Kulkarni, Upasana A1 - Fähnrich, Anke A1 - Bommert, Kurt A1 - Bargou, Ralf A1 - Berek, Claudia A1 - Van, Trung Chu A1 - Bogen, Bjarne A1 - Jundt, Franziska A1 - Manz, Rudolf Armin T1 - Eosinophils and Megakaryocytes Support the Early Growth of Murine MOPC315 Myeloma Cells in Their Bone Marrow Niches JF - PLOS ONE N2 - Multiple myeloma is a bone marrow plasma cell tumor which is supported by the external growth factors APRIL and IL-6, among others. Recently, we identified eosinophils and megakaryocytes to be functional components of the micro-environmental niches of benign bone marrow plasma cells and to be important local sources of these cytokines. Here, we investigated whether eosinophils and megakaryocytes also support the growth of tumor plasma cells in the MOPC315. BM model for multiple myeloma. As it was shown for benign plasma cells and multiple myeloma cells, IL-6 and APRIL also supported MOPC315. BM cell growth in vitro, IL-5 had no effect. Depletion of eosinophils in vivo by IL-5 blockade led to a reduction of the early myeloma load. Consistent with this, myeloma growth in early stages was retarded in eosinophil-deficient Delta dblGATA-1 mice. Late myeloma stages were unaffected, possibly due to megakaryocytes compensating for the loss of eosinophils, since megakaryocytes were found to be in contact with myeloma cells in vivo and supported myeloma growth in vitro. We conclude that eosinophils and megakaryocytes in the niches for benign bone marrow plasma cells support the growth of malignant plasma cells. Further investigations are required to test whether perturbation of these niches represents a potential strategy for the treatment of multiple myeloma. KW - plasma cells KW - human multiple-myeloma KW - immune response KW - receptor expression KW - B-cells KW - stromal cells KW - dexamethasone KW - april KW - survival KW - Interleukin-5 Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-115269 VL - 9 IS - 10 ER - TY - JOUR A1 - Morton, Charles Oliver A1 - Fliesser, Mirjam A1 - Dittrich, Marcus A1 - Müller, Tobias A1 - Bauer, Ruth A1 - Kneitz, Susanne A1 - Hope, William A1 - Rogers, Thomas Richard A1 - Einsele, Hermann A1 - Löffler, Jürgen T1 - Gene Expression Profiles of Human Dendritic Cells Interacting with Aspergillus fumigatus in a Bilayer Model of the Alveolar Epithelium/Endothelium Interface N2 - The initial stages of the interaction between the host and Aspergillus fumigatus at the alveolar surface of the human lung are critical in the establishment of aspergillosis. Using an in vitro bilayer model of the alveolus, including both the epithelium (human lung adenocarcinoma epithelial cell line, A549) and endothelium (human pulmonary artery epithelial cells, HPAEC) on transwell membranes, it was possible to closely replicate the in vivo conditions. Two distinct sub-groups of dendritic cells (DC), monocyte-derived DC (moDC) and myeloid DC (mDC), were included in the model to examine immune responses to fungal infection at the alveolar surface. RNA in high quantity and quality was extracted from the cell layers on the transwell membrane to allow gene expression analysis using tailored custom-made microarrays, containing probes for 117 immune-relevant genes. This microarray data indicated minimal induction of immune gene expression in A549 alveolar epithelial cells in response to germ tubes of A. fumigatus. In contrast, the addition of DC to the system greatly increased the number of differentially expressed immune genes. moDC exhibited increased expression of genes including CLEC7A, CD209 and CCL18 in the absence of A. fumigatus compared to mDC. In the presence of A. fumigatus, both DC subgroups exhibited up-regulation of genes identified in previous studies as being associated with the exposure of DC to A. fumigatus and exhibiting chemotactic properties for neutrophils, including CXCL2, CXCL5, CCL20, and IL1B. This model closely approximated the human alveolus allowing for an analysis of the host pathogen interface that complements existing animal models of IA. KW - aspergillus fumigatus KW - gene expression KW - immune receptors KW - immune response KW - denritic cells KW - B cell receptors KW - gene regulation KW - RNA extraction Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-112893 ER - TY - JOUR A1 - Ronchi, Cristina L. A1 - Sbiera, Silviu A1 - Volante, Marco A1 - Steinhauer, Sonja A1 - Scott-Wild, Vanessa A1 - Altieri, Barbara A1 - Kroiss, Matthias A1 - Bala, Margarita A1 - Papotti, Mauro A1 - Deutschbein, Timo A1 - Terzolo, Massimo A1 - Fassnacht, Martin A1 - Allolio, Bruno T1 - CYP2W1 Is Highly Expressed in Adrenal Glands and Is Positively Associated with the Response to Mitotane in Adrenocortical Carcinoma N2 - Background Adrenocortical tumors comprise frequent adenomas (ACA) and rare carcinomas (ACC). Human cytochrome P450 2W1 (CYP2W1) is highly expressed in some cancers holding the potential to activate certain drugs into tumor cytotoxins. Objective To investigate the CYP2W1 expression in adrenal samples and its relationship with clinical outcome in ACC. Material and Methods CYP2W1 expression was investigated by qRT-PCR in 13 normal adrenal glands, 32 ACA, 25 ACC, and 9 different non-adrenal normal tissue samples and by immunohistochemistry in 352 specimens (23 normal adrenal glands, 33 ACA, 239 ACC, 67 non-adrenal normal or neoplastic samples). Results CYP2W1 mRNA expression was absent/low in normal non-adrenal tissues, but high in normal and neoplastic adrenal glands (all P<0.01 vs non-adrenal normal tissues). Accordingly, CYP2W1 immunoreactivity was absent/low (H-score 0–1) in 72% of non-adrenal normal tissues, but high (H-score 2–3) in 44% of non-adrenal cancers, in 65% of normal adrenal glands, in 62% of ACAs and in 50% of ACCs (all P<0.001 vs non-adrenal normal tissues), being significantly increased in steroid-secreting compared to non-secreting tumors. In ACC patients treated with mitotane only, high CYP2W1 immunoreactivity adjusted for ENSAT stage was associated with longer overall survival and time to progression (P<0.05 and P<0.01, respectively), and with a better response to therapy both as palliative (response/stable disease in 42% vs 6%, P<0.01) or adjuvant option (absence of disease recurrence in 69% vs 45%, P<0.01). Conclusion CYP2W1 is highly expressed in both normal and neoplastic adrenal glands making it a promising tool for targeted therapy in ACC. Furthermore, CYP2W1 may represent a new predictive marker for the response to mitotane treatment. KW - CYP2W1 KW - cancer treatment KW - adrenal glands KW - carcinomas KW - drug therapy KW - hormones KW - immune response KW - immunohistochemistry techniques KW - surgical oncology Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-113096 ER - TY - JOUR A1 - Ardelt, Peter U. A1 - Ebbing, Jan A1 - Adams, Fabian A1 - Reiss, Cora A1 - Arap, Wadih A1 - Pasqualini, Renata A1 - Bachmann, Alexander A1 - Wetterauer, Ulrich A1 - Riedmiller, Hubertus A1 - Kneitz, Burkard T1 - An anti-ubiquitin antibody response in transitional cell carcinoma of the urinary bladder JF - PLoS ONE N2 - Background To use combinatorial epitope mapping ("fingerprinting") of the antibody response to identify targets of the humoral immune response in patients with transitional cell carcinoma (TCC) of the bladder. Methods A combinatorial random peptide library was screened on the circulating pool of immunoglobulins purified from an index patient with a high risk TCC (pTa high grade plus carcinoma in situ) to identify corresponding target antigens. A patient cohort was investigated for antibody titers against ubiquitin. Results We selected, isolated, and validated an immunogenic peptide motif from ubiquitin as a dominant epitope of the humoral response. Patients with TCC had significantly higher antibody titers against ubiquitin than healthy donors (p<0.007), prostate cancer patients (p<0.0007), and all patients without TCC taken together (p<0.0001). Titers from superficial tumors were not significantly different from muscle invasive tumors (p = 0.0929). For antibody response against ubiquitin, sensitivity for detection of TCC was 0.44, specificity 0.96, positive predictive value 0.96 and negative predictive value 0.41. No significant titer changes were observed during the standard BCG induction immunotherapy. Conclusions This is the first report to demonstrate an anti-ubiquitin antibody response in patients with TCC. Although sensitivity of antibody production was low, a high specificity and positive predictive value make ubiquitin an interesting candidate for further diagnostic and possibly immune modulating studies. KW - Bacillus-Calmette-Guerin KW - immune response KW - ubiquitin KW - protein biomarkers KW - system bcg KW - tumor cells KW - immunotherapy KW - cancer surveillance Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-143711 VL - 10 IS - 3 ER -