@article{BoeckelKarstenGoepeletal.2023, author = {Boeckel, Hannah and Karsten, Christian M. and G{\"o}pel, Wolfgang and Herting, Egbert and Rupp, Jan and H{\"a}rtel, Christoph and Hartz, Annika}, title = {Increased expression of anaphylatoxin C5a-receptor-1 in neutrophils and natural killer cells of preterm infants}, series = {International Journal of Molecular Sciences}, volume = {24}, journal = {International Journal of Molecular Sciences}, number = {12}, issn = {1422-0067}, doi = {10.3390/ijms241210321}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-321196}, year = {2023}, abstract = {Preterm infants are susceptible to infection and their defense against pathogens relies largely on innate immunity. The role of the complement system for the immunological vulnerability of preterm infants is less understood. Anaphylatoxin C5a and its receptors C5aR1 and -2 are known to be involved in sepsis pathogenesis, with C5aR1 mainly exerting pro-inflammatory effects. Our explorative study aimed to determine age-dependent changes in the expression of C5aR1 and C5aR2 in neonatal immune cell subsets. Via flow cytometry, we analyzed the expression pattern of C5a receptors on immune cells isolated from peripheral blood of preterm infants (n = 32) compared to those of their mothers (n = 25). Term infants and healthy adults served as controls. Preterm infants had a higher intracellular expression of C5aR1 on neutrophils than control individuals. We also found a higher expression of C5aR1 on NK cells, particularly on the cytotoxic CD56\(^{dim}\) subset and the CD56\(^-\) subset. Immune phenotyping of other leukocyte subpopulations revealed no gestational-age-related differences for the expression of and C5aR2. Elevated expression of C5aR1 on neutrophils and NK cells in preterm infants may contribute to the phenomenon of "immunoparalysis" caused by complement activation or to sustained hyper-inflammatory states. Further functional analyses are needed to elucidate the underlying mechanisms.}, language = {en} } @phdthesis{Krenz2023, author = {Krenz, Bastian}, title = {The immune-evasive potential of MYC in pancreatic ductal adenocarcinoma}, doi = {10.25972/OPUS-32590}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-325903}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2023}, abstract = {Pancreatic ductal adenocarcinoma (PDAC) is predominantly driven by mutations in KRAS and TP53. However, PDAC tumors display deregulated levels of MYC and are a paradigm example for MYC-driven and -addicted tumors. For many years MYC was described as a transcription factor that regulates a pleiotropic number of genes to drive proliferation. Recent work sheds a different light on MYC biology. First, changes in gene expression that come along with the activation of MYC are mild and MYC seems to act more as a factor that reduces stress and increases resilience towards challenges during transcription. Second, MYC is a strong driver of immune evasion in different entities. In this study we depleted MYC in murine PDAC cells and revealed the immune dependent regression of tumors in an orthotope transplant model, as well as the activation of the innate immune system using global expression analysis, immunoblotting and fCLIP. These experiments revealed that endogenous double-stranded RNA is binding as a viral mimicry to Toll-like receptor 3, causing activation of TBK1 and downstream activation of a proimmunogenic transcription program. The regression of tumors upon depletion of MYC is dependent on this pathway since the knockout of TBK1 prevents regression of tumors after depletion of MYC. We can summarize this study in three main findings: First, the dominant and most important function of MYC in tumors is not to drive proliferation but to promote immune evasion and prevent immune-dependent regression of tumors. Second, cells monitor defects or delay in splicing and RNA processing and activate the immune system to clear cells that face problems with co-transcriptional processing. Third, MYC suppresses the activation of the cell-intrinsic innate immune system and shields highly proliferating cells from the recognition by the immune system. To translate this into a therapeutically approach, we replaced the shRNA mediated depletion of MYC by treatment with cardiac glycosides. Upon treatment with cardiac glycosides tumor cells reduce uptake of nutrients, causing a downregulation of MYC translation, inhibition of proliferation, glycolysis and lactate secretion. Lactate is a major reason for immune evasion in solid tumors since it dampens, amongst others, cytotoxic T cells and promotes regulatory T cells. Treatment of mice with cardiac glycosides causes a complete and immune-dependent remission of PDAC tumors in vivo, pointing out that cardiac glycosides have strong proimmunogenic, anti-cancer effects. More detailed analyses will be needed to dissect the full mechanism how cardiac glycosides act on MYC translation and immune evasion in PDAC tumors.}, subject = {Bauchspeicheldr{\"u}senkrebs}, language = {en} } @article{HergovitsMaisHaanetal.2017, author = {Hergovits, Sabine and Mais, Christine and Haan, Claude and Costa-Pereira, Ana P. and Hermanns, Heike M.}, title = {Oncostatin M induces RIG-I and MDA5 expression and enhances the double-stranded RNA response in fibroblasts}, series = {Journal of Cellular and Molecular Medicine}, volume = {21}, journal = {Journal of Cellular and Molecular Medicine}, number = {11}, doi = {10.1111/jcmm.13221}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-159558}, pages = {3087-3099}, year = {2017}, abstract = {Interleukin (IL)-6-type cytokines have no direct antiviral activity; nevertheless, they display immune-modulatory functions. Oncostatin M (OSM), a member of the IL-6 family, has recently been shown to induce a distinct number of classical interferon stimulated genes (ISG). Most of them are involved in antigen processing and presentation. However, induction of retinoic acid-inducible gene (RIG)-I-like receptors (RLR) has not been investigated. Here we report that OSM has the capability to induce the expression of the DExD/H-Box RNA helicases RIG-I and melanoma differentiation antigen 5 (MDA5) as well as of the transcription factors interferon regulatory factor (IRF)1, IRF7 and IRF9 in primary fibroblasts. Induction of the helicases depends on tyrosine as well as serine phosphorylation of STAT1. Moreover, we could show that the OSM-induced STAT1 phosphorylation is predominantly counter-regulated by a strong STAT3-dependent SOCS3 induction, as Stat3 as well as Socs3 knock-down results in an enhanced and prolonged helicase and IRF expression. Other factors involved in regulation of STAT1 or IRF1 activity, like protein tyrosine phosphatase, non-receptor type 2 (PTPN2), promyelocytic leukaemia protein (PML) or small ubiquitin-related modifier 1 (SUMO1), play a minor role in OSM-mediated induction of RLR. Remarkably, OSM and interferon-γ (IFN-γ) synergize to mediate transcription of RLR and pre-treatment of fibroblasts with OSM fosters the type I interferon production in response to a subsequent encounter with double-stranded RNA. Together, these findings suggest that the OSM-induced JAK/STAT1 signalling is implicated in virus protection of non-professional immune cells and may cooperate with interferons to enhance RLR expression in these cells.}, language = {en} } @article{SagivMichaeliAssietal.2015, author = {Sagiv, Jitka Y. and Michaeli, Janna and Assi, Simaan and Mishalian, Inbal and Kisos, Hen and Levy, Liran and Damti, Pazzit and Lumbroso, Delphine and Polyansky, Lola and Sionov, Ronit V. and Ariel, Amiram and Hovav, Avi-Hai and Henke, Erik and Fridlender, Zvi G. and Granot, Zvi}, title = {Phenotypic diversity and plasticity in circulating neutrophil subpopulations in cancer}, series = {Cell Reports}, volume = {10}, journal = {Cell Reports}, number = {4}, doi = {10.1016/j.celrep.2014.12.039}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-144102}, pages = {562-573}, year = {2015}, abstract = {Controversy surrounds neutrophil function in cancer because neutrophils were shown to provide both pro-and antitumor functions. We identified a heterogeneous subset of low-density neutrophils (LDNs) that appear transiently in self-resolving inflammation but accumulate continuously with cancer progression. LDNs display impaired neutrophil function and immunosuppressive properties, characteristics that are in stark contrast to those of mature, high-density neutrophils (HDNs). LDNs consist of both immature myeloid-derived suppressor cells (MDSCs) and mature cells that are derived from HDNs in a TGF-beta-dependent mechanism. Our findings identify three distinct populations of circulating neutrophils and challenge the concept that mature neutrophils have limited plasticity. Furthermore, our findings provide a mechanistic explanation to mitigate the controversy surrounding neutrophil function in cancer.}, language = {en} } @article{LoefflerLoefflerKobsaretal.2015, author = {Loeffler, Claudia and Loeffler, J{\"u}rgen and Kobsar, Anna and Speer, Christian P. and Eigenthaler, Martin}, title = {Septic Vs Colonizing Group B Streptococci Differentially Regulate Inflammation and Apoptosis in Human Coronary Artery Endothelial Cells - a Pilot Study}, series = {Journal of Pediatrics and Neonatal Care}, volume = {2}, journal = {Journal of Pediatrics and Neonatal Care}, number = {2}, doi = {10.15406/jpnc.2015.02.00064}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-125596}, pages = {00064}, year = {2015}, abstract = {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.}, language = {en} } @article{WilhelmSmetakSchaeferEckartetal.2014, author = {Wilhelm, Martin and Smetak, Manfred and Schaefer-Eckart, Kerstin and Kimmel, Brigitte and Birkmann, Josef and Einsele, Hermann and Kunzmann, Volker}, title = {Successful adoptive transfer and in vivo expansion of haploidentical γδ T cells}, series = {Journal of Translational Medicine}, volume = {12}, journal = {Journal of Translational Medicine}, number = {45}, doi = {10.1186/1479-5876-12-45}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-117290}, year = {2014}, abstract = {Background: The primary aim of this pilot study was to determine the feasibility and safety of an adoptive transfer and in vivo expansion of human haploidentical gamma delta T lymphocytes. Methods: Patients with advanced haematological malignancies who are not eligible for allogeneic transplantation received peripheral blood mononuclear cells from half-matched family donors. For that, a single unstimulated leukapheresis product was incubated with both the anti-CD4 and anti-CD8 antibodies conjugated to paramagnetic particles. The depletion procedure was performed on a fully automated CliniMACS (R) device according to the manufacturer's instructions. On average, patients received 2.17 x 10(6)/kg (range 0.9-3.48) γδ T cells with <1\% CD4-or CD8-positive cells remaining in the product. All patients received prior lymphopenia-inducing chemotherapy (fludarabine 20-25 mg/m(2) day -6 until day -2 and cyclophosphamide 30-60 mg/kg day -6 and -5) and were treated with 4 mg zoledronate on day 0 and 1.0x10(6) IU/m(2) IL-2 on day +1 until day +6 for the induction of gamma delta T cell proliferation in vivo. Results: This resulted in a marked in vivo expansion of donor γδ T cells and, to a lower extent, natural killer cells and double-negative αβ T cells (mean 68-fold, eight-fold, and eight-fold, respectively). Proliferation peaked by around day +8 and donor cells persisted up to 28 days. Although refractory to all prior therapies, three out of four patients achieved a complete remission, which lasted for 8 months in a patient with plasma cell leukaemia. One patient died from an infection 6 weeks after treatment. Conclusion: This pilot study shows that adoptive transfer and in vivo expansion of haploidentical γδ T lymphocytes is feasible and suggests a potential role of these cells in the treatment of haematological diseases.}, language = {en} } @article{GowdaGodderKmieciaketal.2011, author = {Gowda, Madhu and Godder, Kamar and Kmieciak, Maciej and Worschech, Andrea and Ascierto, Maria-Libera and Wang, Ena and Francesco M., Marincola and Manjili, Masoud H.}, title = {Distinct signatures of the immune responses in low risk versus high risk neuroblastoma}, series = {Journal of Translational Medicine}, volume = {9}, journal = {Journal of Translational Medicine}, number = {170}, doi = {10.1186/1479-5876-9-170}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-135147}, pages = {1-8}, year = {2011}, abstract = {Background: Over 90\% of low risk (LR) neuroblastoma patients survive whereas less than 30\% of high risk (HR) patients are long term survivors. Age (children younger than 18 months old) is associated with LR disease. Considering that adaptive immune system is well developed in older children, and that T cells were shown to be involved in tumor escape and progression of cancers, we sought to determine whether HR patients may tend to show a signature of adaptive immune responses compared to LR patients who tend to have diminished T-cell responses but an intact innate immune response. Methods: We performed microarray analysis of RNA extracted from the tumor specimens of HR and LR patients. Flow cytometry was performed to determine the cellular constituents in the blood while multiplex cytokine array was used to detect the cytokine profile in patients' sera. A HR tumor cell line, SK-N-SH, was also used for detecting the response to IL-1 beta, a cytokines which is involved in the innate immune responses. Results: Distinct patterns of gene expression were detected in HR and LR patients indicating an active T-cell response and a diminished adaptive immune response, respectively. A diminished adaptive immune response in LR patients was evident by higher levels of IL-10 in the sera. In addition, HR patients had lower levels of circulating myeloid derived suppressor cells (MDSC) compared with a control LR patient. LR patients showed slightly higher levels of cytokines of the innate immune responses. Treatment of the HR tumor line with IL-1b induced expression of cytokines of the innate immune responses. Conclusions: This data suggests that adaptive immune responses may play an important role in the progression of HR disease whereas innate immune responses may be active in LR patients.}, language = {en} } @phdthesis{Boehme2009, author = {B{\"o}hme, Linda}, title = {Cellular response to double-stranded RNA in Chlamydia trachomatis-infected human host cells}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-46474}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2009}, abstract = {Chlamydien sind Gram-negative, obligat-intrazellul{\"a}re Bakterien, die f{\"u}r ein weites Spektrum an relevanten Krankheiten verantwortlich sind. Auf Grund ihres zweiphasigen Entwicklungszyklusses sind Chlamydien von einer intakten Wirtszelle abh{\"a}ngig, um sich erfolgreich vermehren und im Organismus ausbreiten zu k{\"o}nnen. Daher haben Chlamydien anspruchsvolle Strategien entwickelt, um das Immunsystem des Wirtes auszuschalten oder den programmierten Zelltod ihrer Wirtszelle zu verhindern. In der vorliegenden Arbeit wurde untersucht, ob eine Infektion mit C. trachomatis einen Einfluss auf die zellul{\"a}re Antwort auf dsRNA nehmen kann. Die Synthese von dsRNA ist ein charakteristisches Merkmal der Replikation von Viren, welche sowohl die Apoptose induzieren als auch das Immunsystem aktivieren kann. Um eine chlamydiale und virale Co-Infektion zu simulieren, wurden Chlamydien-infizierte Epithelzellen mit der synthetischen dsRNA Polyinosin-Polycytidins{\"a}ure (polyI:C) transfiziert. Im ersten Teil der Arbeit wurde untersucht, ob Chlamydien die durch dsRNA eingeleitete Apoptose verhindern k{\"o}nnen. Eine signifikante Reduktion der dsRNA-induzierten Apoptose konnte in infizierten Zellen beobachtet werden. Es zeigte sich, dass die Prozessierung der Initiator-Caspase-8 in infizierten Zellen unterblieb. Dies war von der fr{\"u}hen bakteriellen Proteinsynthese abh{\"a}ngig und f{\"u}r die dsRNA-vermittelte Apoptose spezifisch, da der durch TNFalpha bewirkte Zelltod nicht auf der Ebene der Caspase-8 verhindert werden konnte. Die Aktivierung von zellul{\"a}ren Faktoren, die bei der Apoptoseinduzierung eine wichtige Rolle spielen, beispielsweise PKR und RNase L, war in infizierten Zellen jedoch unver{\"a}ndert. Stattdessen konnte durch RNA Interferenz-vermittelte Depletion gezeigt werden, dass der zellul{\"a}re Caspase-8-Inhibitor cFlip eine entscheidende Rolle bei der chlamydialen Blockierung der dsRNA-vermittelten Apoptose spielt. Mittels Co-Immunopr{\"a}zipitation konnte ein erster Hinweis darauf gefunden werden, dass C. trachomatis eine Anreicherung von cFlip im dsRNA-induzierten Komplex von Caspase-8 und FADD bewirkt. Im zweiten Teil der Arbeit wurde untersucht, ob Chlamydien die Immunantwort auf virale Infektionen beeinflussen, welche vor allem die Expression von Interferonen und Interleukinen beinhaltet. Es stellte sich heraus, dass die Aktivierung des Interferon regulatory factor 3 (IRF-3) und des zur Familie von NF-kappaB Trankriptionsfaktoren geh{\"o}renden p65, zwei zentralen Regulatoren der Immunantwort auf dsRNA, in infizierten Epithelzellen ver{\"a}ndert war. Die Degradation von IkappaB-alpha, des Inhibitors von NF-kappaB, war in infizierten Zellen beschleunigt, begleitet von einer Ver{\"a}nderung der Translokation des Transkriptionsfaktors in den Zellkern. Im Gegensatz dazu wurde die nukle{\"a}re Translokation von IRF-3 durch die Infektion signifikant verhindert. Die hier vorgestellten Daten zeigen erstmals, dass eine Infektion mit C. trachomatis die zellul{\"a}re Antwort auf dsRNA signifikant ver{\"a}ndern kann und implizieren einen Einfluss von chlamydialen Infektionen auf den Ausgang von viralen Superinfektionen.}, subject = {Chlamydia trachomatis}, language = {en} }