TY - JOUR A1 - Hausmann, Stefan A1 - Brandt, Evelyn A1 - Köchel, Carolin A1 - Einsele, Hermann A1 - Bargou, Ralf C. A1 - Seggewiss-Bernhardt, Ruth A1 - Stühmer, Thorsten T1 - Loss of serum and glucocorticoid-regulated kinase 3 (SGK3) does not affect proliferation and survival of multiple myeloma cell lines JF - PLoS ONE N2 - Multiple myeloma (MM) is a generally fatal plasma cell cancer that often shows activation of the phosphoinositide 3-kinase/Akt (PI3K/Akt) pathway. Targeted pharmacologic therapies, however, have not yet progressed beyond the clinical trial stage, and given the complexity of the PI3K/Akt signalling system (e.g. multiple protein isoforms, diverse feedback regulation mechanisms, strong variability between patients) it is mandatory to characterise its ramifications in order to better guide informed decisions about the best therapeutic approaches. Here we explore whether serum and glucocorticoid-regulated kinase 3 (SGK3), a potential downstream effector of PI3K, plays a role in oncogenic signalling in MM cells-either in concert with or independent of Akt. SGK3 was expressed in all MM cell lines and in all primary MM samples tested. Four MM cell lines representing a broad range of intrinsic Akt activation (very strong: MM. 1s, moderate: L 363 and JJN-3, absent: AMO-1) were chosen to test the effects of transient SGK3 knockdown alone and in combination with pharmacological inhibition of Akt, PI3K-p110\(\alpha\), or in the context of serum starvation. Although the electroporation protocol led to strong SGK3 depletion for at least 5 days its absence had no substantial effect on the activation status of potential downstream substrates, or on the survival, viability or proliferation of MM cells in all experimental contexts tested. We conclude that it is unlikely that SGK3 plays a significant role for oncogenic signalling in multiple myeloma. KW - Akt KW - phosphorylation KW - downstream KW - mechanism KW - pathway KW - isoforms KW - activation KW - cancer KW - inhibition KW - phosphatidylinositol 3-kinase/Akt Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-148708 VL - 10 IS - 4 ER - TY - JOUR A1 - Becker, Philip P. A1 - Rau, Monika A1 - Schmitt, Johannes A1 - Malsch, Carolin A1 - Hammer, Christian A1 - Bantel, Heike A1 - Müllhaupt, Beat A1 - Geier, Andreas T1 - Performance of serum microRNAs -122, -192 and -21 as biomarkers in patients with non-alcoholic steatohepatitis JF - PLoS ONE N2 - Objectives Liver biopsies are the current gold standard in non-alcoholic steatohepatitis (NASH) diagnosis. Their invasive nature, however, still carries an increased risk for patients' health. The development of non-invasive diagnostic tools to differentiate between bland steatosis (NAFL) and NASH remains crucial. The aim of this study is the evaluation of investigated circulating microRNAs in combination with new targets in order to optimize the discrimination of NASH patients by non-invasive serum biomarkers. Methods Serum profiles of four microRNAs were evaluated in two cohorts consisting of 137 NAFLD patients and 61 healthy controls. In a binary logistic regression model microRNAs of relevance were detected. Correlation of microRNA appearance with known biomarkers like ALT and CK18-Asp396 was evaluated. A simplified scoring model was developed, combining the levels of microRNA in circulation and CK18-Asp396 fragments. Receiver operating characteristics were used to evaluate the potential of discriminating NASH. Results The new finding of our study is the different profile of circulating miR-21 in NASH patients (p<0.0001). Also, it validates recently published results of miR-122 and miR-192 to be differentially regulated in NAFL and NASH. Combined microRNA expression profiles with CK18-Asp396 fragment level scoring model had a higher potential of NASH prediction compared to other risk biomarkers (AUROC = 0.83, 95% CI = 0.754-0.908; p<0.001). Evaluation of score model for NAFL (Score = 0) and NASH (Score = 4) had shown high rates of sensitivity (91%) and specificity (83%). Conclusions Our study defines candidates for a combined model of miRNAs and CK18-Asp396 levels relevant as a promising expansion for diagnosis and in turn treatment of NASH. KW - fatty liver disease KW - independent marker KW - expression KW - injury KW - NAFLD KW - circulating micrornas KW - caspase activation KW - fibrosis KW - miR-122 KW - apoptosis Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-145147 VL - 10 IS - 11 ER - TY - JOUR A1 - Barej, Michael F. A1 - Schmitz, Andreas A1 - Penner, Johannes A1 - Doumbia, Joseph A1 - Sandberger-Loua, Laura A1 - Hirschfeld, Mareike A1 - Brede, Christian A1 - Emmrich, Mike A1 - Kouamé, N'Goran Germain A1 - Hillers, Annika A1 - Gonwouo, Nono L. A1 - Nopper, Joachim A1 - Adeba, Patrick Joël A1 - Bangoura, Mohamed A. A1 - Gage, Ceri A1 - Anderson, Gail A1 - Rödel, Mark-Oliver T1 - Life in the spray zone - overlooked diversity in West African torrent-frogs (Anura, Odontobatrachidae, Odontobatrachus) JF - Zoosystematics and Evolution N2 - West African torrent-frogs of the genus Odontobatrachus currently belong to a single species: Odontobatrachus natator (Boulenger, 1905). Recently, molecular results and biogeographic separation led to the recognition of five Operational Taxonomic Units (OTUs) thus identifying a species-complex. Based on these insights, morphological analyses on more than 150 adult specimens, covering the entire distribution of the family and all OTUs, were carried out. Despite strong morphological congruence, combinations of morphological characters made the differentiation of OTUs successful and allowed the recognition of five distinct species: Odontobatrachus natator, and four species new to science: Odontobatrachus arndti sp. n., O. fouta sp. n., O. smithi sp. n. and O. ziama sp. n. All species occur in parapatry: Odontobatrachus natator is known from western Guinea to eastern Liberia, O. ziama sp. n. from eastern Guinea, O. smithi sp. n. and O. fouta sp. n. from western Guinea, O. arndti sp. n. from the border triangle Guinea-Liberia-Cote d'Ivoire. In addition, for the first time the advertisement call of a West African torrent-frog (O. arndti sp. n.) is described. KW - Guinean rain forest KW - molecular data KW - conservation KW - Upper Guinea KW - new species KW - Phrynobatrachus amphibia KW - Arthroleptis amphibia KW - ivory coast KW - genus KW - biodiversity KW - Ranidae KW - Petropedetidae KW - biodiversity hotspot KW - rainforest KW - taxonomy KW - Amphibia Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-144254 VL - 91 IS - 2 ER - TY - JOUR A1 - Andersen, Jens Peter A1 - Bøgsted, Martin A1 - Dybkær, Karen A1 - Mellqvist, Ulf-Henrik A1 - Morgan, Gareth J. A1 - Goldschmidt, Hartmut A1 - Dimopoulos, Meletios A. A1 - Einsele, Hermann A1 - San Miguel, Jesús A1 - Palumbo, Antonio A1 - Sonneveld, Pieter A1 - Johnsen, Hans Erik T1 - Global myeloma research clusters, output, and citations: a bibliometric mapping and clustering analysis JF - PLoS ONE N2 - Background International collaborative research is a mechanism for improving the development of disease-specific therapies and for improving health at the population level. However, limited data are available to assess the trends in research output related to orphan diseases. Methods and Findings We used bibliometric mapping and clustering methods to illustrate the level of fragmentation in myeloma research and the development of collaborative efforts. Publication data from Thomson Reuters Web of Science were retrieved for 2005-2009 and followed until 2013. We created a database of multiple myeloma publications, and we analysed impact and co-authorship density to identify scientific collaborations, developments, and international key players over time. The global annual publication volume for studies on multiple myeloma increased from 1,144 in 2005 to 1,628 in 2009, which represents a 43% increase. This increase is high compared to the 24% and 14% increases observed for lymphoma and leukaemia. The major proportion (> 90% of publications) was from the US and EU over the study period. The output and impact in terms of citations, identified several successful groups with a large number of intra-cluster collaborations in the US and EU. The US-based myeloma clusters clearly stand out as the most productive and highly cited, and the European Myeloma Network members exhibited a doubling of collaborative publications from 2005 to 2009, still increasing up to 2013. Conclusion and Perspective Multiple myeloma research output has increased substantially in the past decade. The fragmented European myeloma research activities based on national or regional groups are progressing, but they require a broad range of targeted research investments to improve multiple myeloma health care. KW - multiparametric flow cytometry KW - multiple myeloma KW - consensus statement KW - European experts KW - disorders KW - therapy KW - network Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-144214 VL - 10 IS - 1 ER - TY - JOUR A1 - Spinner, Christoph D A1 - Wille, Florian A1 - Schwerdtfeger, Christiane A1 - Thies, Philipp A1 - Tanase, Ursula A1 - Von Figura, Guido A1 - Schmid, Roland M A1 - Heinz, Werner J A1 - Klinker, Hartwig Hf T1 - Pharmacokinetics of chewed vs. swallowed raltegravir in a patient with AIDS and MAI infection: some new conflicting data JF - AIDS Research and Therapy N2 - Background: While HIV, AIDS and atypical Mycobacterium infections are closely linked, the use of Integrase-Inhibitor based cART, notably raltegravir-based regimens is more widespread. RAL should be double-dosed to 800 mg semi-daily in situation of rifampicin co-medication, because RAL is more rapidly metabolized due to rifampicin-induced Uridine-5'-diphosph-gluronosyl-transferase (UGT1A1). Recently, it was speculated that chewed RAL might lead to increased absorption, which might compensate the inductive effect of rifampicin-rapid metabolized RAL, as part of cost-saving effects in countries with high-tuberculosis prevalence and less economic power. Methods: We report measurement of raltegravir pharmacokinetics in a 34-year AIDS-patient suffering from disseminated Mycobacterium avium infection with necessity of parenteral rifampicin treatment. RAL levels were measured with HPLC (internal standard: carbamazepine, LLQ 11 ng/ml, validation with Valistat 2.0 program (Arvecon, Germany)). For statistical analysis, a two-sided Wilcoxon signed rank test for paired samples was used. Results: High intra-personal variability in raltegravir serum levels was seen. Comparable C\(_{max}\) concentrations were found for 800 mg chewed and swallowed RAL, as well as for 400 mg chewed and swallowed RAL. While C\(_{max}\) seems to be more dependent from overall RAL dosing than from swallowed or chewed tablets, increased AUC(12) is clearly linked to higher RAL dosages per administration. Anyway, chewed raltegravir showed a rapid decrease in serum levels. Conclusions: We found no evidence that chewed 400 mg semi-daily raltegravir in rifampicin co-medication leads to optimized pharmacokinetics. There is need for more data from randomized trials for further recommendations. KW - pharmacology KW - drug KW - HIV KW - chewed KW - Mycobacterium avium KW - raltegravir KW - pharmacokinetic Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-144058 VL - 12 IS - 1 ER - TY - JOUR A1 - Leonhardt, Ines A1 - Spielberg, Steffi A1 - Weber, Michael A1 - Albrecht-Eckardt, Daniela A1 - Bläss, Markus A1 - Claus, Ralf A1 - Barz, Dagmar A1 - Scherlach, Kirstin A1 - Hertweck, Christian A1 - Löffler, Jürgen A1 - Hünniger, Kerstin A1 - Kurzai, Oliver T1 - The fungal quorum-sensing molecule farnesol activates innate immune cells but suppresses cellular adaptive immunity JF - mBio N2 - Farnesol, produced by the polymorphic fungus Candida albicans, is the first quorum-sensing molecule discovered in eukaryotes. Its main function is control of C. albicans filamentation, a process closely linked to pathogenesis. In this study, we analyzed the effects of farnesol on innate immune cells known to be important for fungal clearance and protective immunity. Farnesol enhanced the expression of activation markers on monocytes (CD86 and HLA-DR) and neutrophils (CD66b and CD11b) and promoted oxidative burst and the release of proinflammatory cytokines (tumor necrosis factor alpha [TNF-\(\alpha\)] and macrophage inflammatory protein 1 alpha [MIP-1 \(\alpha\)]). However, this activation did not result in enhanced fungal uptake or killing. Furthermore, the differentiation of monocytes to immature dendritic cells (iDC) was significantly affected by farnesol. Several markers important for maturation and antigen presentation like CD1a, CD83, CD86, and CD80 were significantly reduced in the presence of farnesol. Furthermore, farnesol modulated migrational behavior and cytokine release and impaired the ability of DC to induce T cell proliferation. Of major importance was the absence of interleukin 12 (IL-12) induction in iDC generated in the presence of farnesol. Transcriptome analyses revealed a farnesol-induced shift in effector molecule expression and a down-regulation of the granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor during monocytes to iDC differentiation. Taken together, our data unveil the ability of farnesol to act as a virulence factor of C. albicans by influencing innate immune cells to promote inflammation and mitigating the Th1 response, which is essential for fungal clearance. KW - human dendritic cells KW - Pseudomonas aeruginosa KW - induced apoptosis KW - cytokine production KW - biofilm formation KW - Candida albicans KW - mouse model KW - systemic candidiasis KW - oxidative stress KW - carcinoma cells Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-143756 VL - 6 IS - 2 ER - TY - JOUR A1 - Kremer, Joel M A1 - Kivitz, Alan J A1 - Simon-Campos, Jesus A A1 - Nasonov, Evgeny L A1 - Tony, Hans-Peter A1 - Lee, Soo-Kon A1 - Vlahos, Bonnie A1 - Hammond, Constance A1 - Bukowski, Jack A1 - Li, Huihua A1 - Schulman, Seth L A1 - Raber, Susan A1 - Zuckerman, Andrea A1 - Isaacs, John D T1 - Evaluation of the effect of tofacitinib on measured glomerular filtration rate in patients with active rheumatoid arthritis: results from a randomised controlled trial JF - Arthritis Research & Therapy N2 - Introduction: Tofacitinib is an oral Janus kinase inhibitor for the treatment of rheumatoid arthritis (RA). During the clinical development programme, increases in mean serum creatinine (SCr) of approximately 0.07 mg/dL and 0.08 mg/dL were observed which plateaued early. This study assessed changes in measured glomerular filtration rate (mGFR) with tofacitinib relative to placebo in patients with active RA. Methods: This was a randomised, placebo-controlled, Phase 1 study (NCT01484561). Patients were aged \(\geq\)18 years with active RA. Patients were randomised 2: 1 to oral tofacitinib 10 mg twice daily (BID) in Period 1 then placebo BID in Period 2 (tofacitinib -> placebo); or oral placebo BID in both Periods (placebo. placebo). Change in mGFR was evaluated by iohexol serum clearance at four time points (run-in, pre-dose in Period 1, Period 1 end, and Period 2 end). The primary endpoint was the change in mGFR from baseline to Period 1 end. Secondary endpoints included: change in mGFR at other time points; change in estimated GFR (eGFR; Cockcroft-Gault equation) and SCr; efficacy; and safety. Results: 148 patients were randomised to tofacitinib -> placebo (N = 97) or placebo -> placebo (N = 51). Baseline characteristics were similar between groups. A reduction of 8% (90% confidence interval [CI]: 2%, 14%) from baseline in adjusted geometric mean mGFR was observed during tofacitinib treatment in Period 1 vs placebo. During Period 2, mean mGFR returned towards baseline during placebo treatment, and there was no difference between the two treatment groups at the end of the study - ratio (tofacitinib -> placebo/placebo -> placebo) of adjusted geometric mean fold change of mGFR was 1.04 (90% CI: 0.97, 1.11). Post-hoc analyses, focussed on mGFR variability in placebo -> placebo patients, were consistent with this conclusion. At study end, similar results were observed for eGFR and SCr. Clinical efficacy and safety were consistent with prior studies. Conclusion: Increases in mean SCr and decreases in eGFR in tofacitinib-treated patients with RA may occur in parallel with decreases in mean mGFR; mGFR returned towards baseline after tofacitinib discontinuation, with no significant difference vs placebo, even after post-hoc analyses. Safety monitoring will continue in ongoing and future clinical studies and routine pharmacovigilance. KW - janus kinase inhibitor KW - renal function KW - CP-690,550 KW - iohexol KW - disease comorbidities KW - plasma clearance KW - serum creatinine KW - kidney function KW - methotrexate Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-143409 VL - 17 IS - 95 ER - TY - JOUR A1 - Lee, Marcel A1 - Eyer, Florian A1 - Felgenhauer, Norbert A1 - Klinker, Hartwig H. F. A1 - Spinner, Christoph D. T1 - Overdose of dolutegravir in combination with tenofovir disaproxil fumarate/emtricitabine in suicide attempt in a 21-year old patient JF - AIDS Research and Therapy N2 - A 21 year old MSM patient with newly diagnosed HIV infection was hospitalized in our department after ingestion of an overdose of his antiretroviral therapy (ART) comprising dolutegravir (DTG - Tivicay\(^{®}\)) and tenofovir disaproxil fumarate/emtricitabine (Truvada\(^{®}\)) in suicidal intention. On admission, the patient did not show any clinical signs of intoxication and laboratory findings were unremarkable. After 6 hours of intensive care monitoring, the patient was referred to a psychiatric clinic. 5 days after the day of intoxication, serum creatinine levels increased to high normal values (1.2 mg/dl). However, levels never exceeded the upper threshold. 8 and 12 weeks later, serum creatinine normalized to levels measured prior to the intoxication. No other adverse events occurred, and the patient does not suffer from permanent impairments. KW - integrase inhibitor KW - HIV KW - AIDS KW - suicide attempt KW - dolutegravir KW - tenofovir disaproxil fumarate KW - emtricitabine KW - overdose Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-151994 VL - 12 IS - 18 ER - TY - JOUR A1 - Heidrich, Benjamin A1 - Cordes, Hans-Jörg A1 - Klinker, Hartwig A1 - Möller, Bernd A1 - Naumann, Uwe A1 - Rössle, Martin A1 - Kraus, Michael R. A1 - Böker, Klaus H. A1 - Roggel, Christoph A1 - Schuchmann, Marcus A1 - Stoehr, Albrecht A1 - Trein, Andreas A1 - Hardtke, Svenja A1 - Gonnermann, Andrea A1 - Koch, Armin A1 - Wedemeyer, Heiner A1 - Manns, Michael P. A1 - Cornberg, Markus T1 - Treatment Extension of Pegylated Interferon Alpha and Ribavirin Does Not Improve SVR in Patients with Genotypes 2/3 without Rapid Virological Response (OPTEX Trial): A Prospective, Randomized, Two-Arm, Multicentre Phase IV Clinical Trial JF - PLoS ONE N2 - Although sofosbuvir has been approved for patients with genotypes 2/3 (G2/3), many parts of the world still consider pegylated Interferon alpha (P) and ribavirin (R) as standard of care for G2/3. Patients with rapid virological response (RVR) show response rates >80%. However, SVR (sustained virological response) in non-RVR patients is not satisfactory. Longer treatment duration may be required but evidence from prospective trials are lacking. A total of 1006 chronic HCV genotype 2/3 patients treated with P/R were recruited into a German HepNet multicenter screening registry. Of those, only 226 patients were still HCV RNA positive at week 4 (non-RVR). Non-RVR patients with ongoing response after 24 weeks P-2b/R qualified for OPTEX, a randomized trial investigating treatment extension of additional 24 weeks (total 48 weeks, Group A) or additional 12 weeks (total 36 weeks, group B) of 1.5 \(\mu\)g/kg P-2b and 800-1400 mg R. Due to the low number of patients without RVR, the number of 150 anticipated study patients was not met and only 99 non-RVR patients (n=50 Group A, n=49 Group B) could be enrolled into the OPTEX trial. Baseline factors did not differ between groups. Sixteen patients had G2 and 83 patients G3. Based on the ITT (intention-to-treat) analysis, 68% [55%; 81%] in Group A and 57% [43%; 71%] in Group B achieved SVR (p=0.31). The primary endpoint of better SVR rates in Group A compared to a historical control group (SVR 70%) was not met. In conclusion, approximately 23% of G2/3 patients did not achieve RVR in a real world setting. However, subsequent recruitment in a treatment-extension study was difficult. Prolonged therapy beyond 24 weeks did not result in higher SVR compared to a historical control group. KW - chronic hepatitis C KW - peginterferon alpha-2b KW - infection KW - sofosbuvir KW - therapy KW - HCV genotype 2 Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-151811 VL - 10 IS - 6 ER - TY - JOUR A1 - Hecht, Markus A1 - Erber, Sonja A1 - Harrer, Thomas A1 - Klinker, Hartwig A1 - Roth, Thomas A1 - Parsch, Hans A1 - Fiebig, Nora A1 - Fietkau, Rainer A1 - Distel, Luitpold V. T1 - Efavirenz Has the Highest Anti-Proliferative Effect of Non-Nucleoside Reverse Transcriptase Inhibitors against Pancreatic Cancer Cells JF - PLoS ONE N2 - Background Cancer prevention and therapy in HIV-1-infected patients will play an important role in future. The non-nucleoside reverse transcriptase inhibitors (NNRTI) Efavirenz and Nevirapine are cytotoxic against cancer cells in vitro. As other NNRTIs have not been studied so far, all clinically used NNRTIs were tested and the in vitro toxic concentrations were compared to drug levels in patients to predict possible anti-cancer effects in vivo. Methods Cytotoxicity was studied by Annexin-V-APC/7AAD staining and flow cytometry in the pancreatic cancer cell lines BxPC-3 and Panc-1 and confirmed by colony formation assays. The 50% effective cytotoxic concentrations (EC50) were calculated and compared to the blood levels in our patients and published data. Results The in vitro EC50 of the different drugs in the BxPC-3 pancreatic cancer cells were: Efavirenz 31.5\(\mu\)mol/l (= 9944ng/ml), Nevirapine 239\(\mu\)mol/l (= 63786ng/ml), Etravirine 89.0\(\mu\)mol/l (= 38740ng/ml), Lersivirine 543\(\mu\)mol/l (= 168523ng/ml), Delavirdine 171\(\mu\)mol/l (= 78072ng/ml), Rilpivirine 24.4\(\mu\)mol/l (= 8941ng/ml). As Efavirenz and Rilpivirine had the highest cytotoxic potential and Nevirapine is frequently used in HIV-1 positive patients, the results of these three drugs were further studied in Panc-1 pancreatic cancer cells and confirmed with colony formation assays. 205 patient blood levels of Efavirenz, 127 of Rilpivirine and 31 of Nevirapine were analyzed. The mean blood level of Efavirenz was 3587ng/ml (range 162-15363ng/ml), of Rilpivirine 144ng/ml (range 0-572ng/ml) and of Nevirapine 4955ng/ml (range 1856-8697ng/ml). Blood levels from our patients and from published data had comparable Efavirenz levels to the in vitro toxic EC50 in about 1 to 5% of all patients. Conclusion All studied NNRTIs were toxic against cancer cells. A low percentage of patients taking Efavirenz reached in vitro cytotoxic blood levels. It can be speculated that in HIV-1 positive patients having high Efavirenz blood levels pancreatic cancer incidence might be reduced. Efavirenz might be a new option in the treatment of cancer. KW - human hepatic cells KW - active antiretroviral therapy KW - differentiated thyroid tumor KW - HIV-infected patients KW - pharmacokinetic interaction KW - LINE-1 retrotransposition KW - HIV-1-infected subjects KW - healthy volunteers KW - prostate cancer KW - i-131 uptake Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-151694 VL - 10 IS - 6 ER - TY - JOUR A1 - Paholcsek, Melinda A1 - Fidler, Gabor A1 - Konya, Jozsef A1 - Rejto, Laszlo A1 - Mehes, Gabor A1 - Bukta, Evelin A1 - Loeffler, Juergen A1 - Biro, Sandor T1 - Combining standard clinical methods with PCR showed improved diagnosis of invasive pulmonary aspergillosis in patients with hematological malignancies and prolonged neutropenia JF - BMC Infectious Diseases N2 - Background: We assessed the diagnostic value of standard clinical methods and combined biomarker testing (galactomannan assay and polymerase chain reaction screening) in a prospective case-control study to detect invasive pulmonary aspergillosis in patients with hematological malignancies and prolonged neutropenia. Methods: In this observational study 162 biomarker analyses were performed on samples from 27 febrile neutropenic episodes. Sera were successively screened for galactomannan antigen and for Aspergillus fumigatus specific nucleic acid targets. Furthermore thoracic computed tomography scanning was performed along with bronchoscopy with lavage when clinically indicated. Patients were retrospectively stratified to define a case-group with "proven" or "probable" invasive pulmonary aspergillosis (25.93 %) and a control-group of patients with no evidence for of invasive pulmonary aspergillosis (74.07 %). In 44.44 % of episodes fever ceased in response to antibiotic treatment (group II). Empirical antifungal therapy was administered for episodes with persistent or relapsing fever (group I). 48.15 % of patients died during the study period. Postmortem histology was pursued in 53.85 % of fatalities. Results: Concordant negative galactomannan and computed tomography supported by a polymerase chain reaction assay were shown to have the highest discriminatory power to exclude invasive pulmonary aspergillosis. Bronchoalveolar lavage was performed in 6 cases of invasive pulmonary aspergillosis and in 15 controls. Although bronchoalveolar lavage proved negative in 93 % of controls it did not detect IPA in 86 % of the cases. Remarkably post mortem histology convincingly supported the presence of Aspergillus hyphae in lung tissue from a single case which had consecutive positive polymerase chain reaction assay results but was misdiagnosed by both computed tomography and consistently negative galactomannan assay results. For the galactomannan enzyme-immunoassay the diagnostic odds ratio was 15.33 and for the polymerase chain reaction assay it was 28.67. According to Cohen's kappa our in-house polymerase chain reaction method showed a fair agreement with the galactomannan immunoassay. Combined analysis of the results from the Aspergillus galactomannan enzyme immunoassay together with those generated by our polymerase chain reaction assay led to no misdiagnoses in the control group. Conclusion: The data from this pilot-study demonstrate that the consideration of standard clinical methods combined with biomarker testing improves the capacity to make early and more accurate diagnostic decisions. KW - whole blood specimens KW - high risk KW - mold disease KW - therapy KW - combination testing KW - real time PCR KW - fungal infections KW - immunocompromised patients KW - prospective feasibility KW - galactomannan KW - epidemiology KW - invasive pulmonary aspergillosis KW - biomarkers KW - acute leukemia KW - neutropenic fever Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-151607 VL - 15 IS - 251 ER - TY - JOUR A1 - Irmer, Henriette A1 - Tarazona, Sonia A1 - Sasse, Christoph A1 - Olbermann, Patrick A1 - Loeffler, Jürgen A1 - Krappmann, Sven A1 - Conesa, Ana A1 - Braus, Gerhard H. T1 - RNAseq analysis of Aspergillus fumigatus in blood reveals a just wait and see resting stage behavior JF - BMC Genomics N2 - Background: Invasive aspergillosis is started after germination of Aspergillus fumigatus conidia that are inhaled by susceptible individuals. Fungal hyphae can grow in the lung through the epithelial tissue and disseminate hematogenously to invade into other organs. Low fungaemia indicates that fungal elements do not reside in the bloodstream for long. Results: We analyzed whether blood represents a hostile environment to which the physiology of A. fumigatus has to adapt. An in vitro model of A. fumigatus infection was established by incubating mycelium in blood. Our model allowed to discern the changes of the gene expression profile of A. fumigatus at various stages of the infection. The majority of described virulence factors that are connected to pulmonary infections appeared not to be activated during the blood phase. Three active processes were identified that presumably help the fungus to survive the blood environment in an advanced phase of the infection: iron homeostasis, secondary metabolism, and the formation of detoxifying enzymes. Conclusions: We propose that A. fumigatus is hardly able to propagate in blood. After an early stage of sensing the environment, virtually all uptake mechanisms and energy-consuming metabolic pathways are shut-down. The fungus appears to adapt by trans-differentiation into a resting mycelial stage. This might reflect the harsh conditions in blood where A. fumigatus cannot take up sufficient nutrients to establish self-defense mechanisms combined with significant growth. KW - Saccharomyces cerevisiae KW - cerebral aspergillosis KW - gene expression KW - Aspergillus fumigatus KW - iron homeostasis KW - invasive pulmonary aspergillosis KW - Candida albicans KW - cell wall KW - lysine biosynthesis KW - human pathogen KW - murine model KW - virulence KW - mRNA-Seq KW - transcriptome KW - human pathogenic fungi KW - secondary metabolite gene cluster KW - detoxification Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-151390 VL - 16 IS - 640 ER - TY - THES A1 - Beitzen-Heineke, Antonia T1 - Invariant Natural Killer T cells possess immune-modulating functions during \(Aspergillus\) \(fumigatus\) infection T1 - Invariante Natürliche Killer T Zellen besitzen immunmodulierende Funktionen in der \(Aspergillus\) \(fumigatus\) Abwehr N2 - Aspergillus fumigatus is the most common cause for invasive fungal infections, a disease associated with high mortality in immune-compromised patients. CD1d-restricted invariant Natural Killer T (iNKT) cells compose a small subset of T cells known to impact the immune response towards various infectious pathogens. To investigate the role of human iNKT cells during A. fumigatus infection, we studied their activation as determined by CD69 expression and cytokine production in response to distinct fungal morphotypes in the presence of different CD1d⁺ antigen presenting cells using flow cytometry and multiplex ELISA. Among CD1d⁺ subpopulations, CD1d⁺CD1c⁺ mDCs showed the highest potential to activate iNKT cells on a per cell basis. The presence of A. fumigatus decreased this effect of CD1d⁺CD1c⁺ mDCs on iNKT cells and led to reduced secretion of TNF-α, G-CSF and RANTES. Production of other Th1 and Th2 cytokines was not affected by the fungus, suggesting an immune-modulating function for human iNKT cells during A. fumigatus infection. N2 - Aspergillus fumigatus ist der häufigste Erreger von invasiven Pilzinfektionen, welche bei immunsupprimierten Patienten mit einer hohen Mortalität einhergehen. CD1d-abhängige invariante Natürliche Killer T (iNKT) Zellen sind eine kleine Subpopulation von T-Zellen, die die Immunantwort auf verschiedene Erreger beeinflussen. Um die Rolle von humanen iNKT Zellen in der Aspergillus fumigatus Abwehr zu untersuchen, wurde in Ko-Kulturen mit iNKT Zellen und CD1d⁺ Antigen präsentierenden Zellen in Anwesenheit von verschiedenen fungalen Morphotypen der Aktivierungsmarker CD69 und die Zytokinproduktion mittels Durchflusszytometrie und Multiplex ELISA untersucht. Unter den CD1d⁺ Subpopulationen wiesen die CD1d⁺CD1c⁺mDCs das höchste aktivierende Potential auf iNKT Zellen auf. Die Anwesenheit von A. fumigatus reduzierte diesen Effekt von CD1d⁺CD1c⁺mDCs auf iNKT Zellen und führte zu einer reduzierten Sekretion von TNF-α, G-CSF und RANTES. Die Produktion von anderen Th1 und Th2 Zytokinen war nicht durch den Pilz beeinflusst. Diese Arbeit suggeriert eine immunmodulierende Funktion von humanen iNKT Zellen in der Abwehr von A. fumigatus. KW - Aspergillus fumigatus KW - T Zelle KW - iNKT KW - CD1d KW - CD1c⁺ mDC Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-144966 ER - TY - JOUR A1 - Dimopoulos, Meletios A. A1 - Weisel, Katja C. A1 - Song, Kevin W. A1 - Delforge, Michel A1 - Karlin, Lionel A1 - Goldschmidt, Hartmut A1 - Moreau, Philippe A1 - Banos, Anne A1 - Oriol, Albert A1 - Garderet, Laurent A1 - Cavo, Michele A1 - Ivanova, Valentina A1 - Alegre, Adrian A1 - Martinez-Lopez, Joaquin A1 - Chen, Christine A1 - Spencer, Andrew A1 - Knop, Stefan A1 - Bahlis, Nizar J. A1 - Renner, Christoph A1 - Yu, Xin A1 - Hong, Kevin A1 - Sternas, Lars A1 - Jacques, Christian A1 - Zaki, Mohamed H. A1 - San Miguel, Jesus F. T1 - Cytogenetics and long-term survival of patients with refractory or relapsed and refractory multiple myeloma treated with pomalidomide and low-dose dexamethasone JF - Haematologica N2 - Patients with refractory or relapsed and refractory multiple myeloma who no longer receive benefit from novel agents have limited treatment options and short expected survival. del(17p) and t(4;14) are correlated with shortened survival. The phase 3 MM-003 trial demonstrated significant progression-free and overall survival benefits from treatment with pomalidomide plus low-dose dexamethasone compared to high-dose dexamethasone among patients in whom bortezomib and lenalidomide treatment had failed. At an updated median follow-up of 15.4 months, the progression-free survival was 4.0 versus 1.9 months (HR, 0.50; P<0.001), and median overall survival was 13.1 versus 8.1 months (HR, 0.72; P=0.009). Pomalidomide plus low-dose dexamethasone, compared with high-dose dexamethasone, improved progression-free survival in patients with del(17p) (4.6 versus 1.1 months; HR, 0.34; P < 0.001), t(4;14) (2.8 versus 1.9 months; HR, 0.49; P=0.028), and in standard-risk patients (4.2 versus 2.3 months; HR, 0.55; P<0.001). Although the majority of patients treated with high-dose dexamethasone took pomalidomide after discontinuation, the overall survival of patients treated with pomalidomide plus low-dose dexamethasone or highdose dexamethasone was 12.6 versus 7.7 months (HR, 0.45; P=0.008) in patients with del(17p), 7.5 versus 4.9 months (HR, 1.12; P=0.761) in those with t(4;14), and 14.0 versus 9.0 months (HR, 0.85; P=0.380) in standard-risk subjects. The overall response rate was higher in patients treated with pomalidomide plus low-dose dexamethasone than in those treated with high-dose dexamethasone both among standard-risk patients (35.2% versus 9.7%) and those with del(17p) (31.8% versus 4.3%), whereas it was similar in patients with t(4; 14) (15.9% versus 13.3%). The safety of pomalidomide plus low-dose dexamethasone was consistent with initial reports. In conclusion, pomalidomide plus low-dose dexamethasone is efficacious in patients with relapsed/refractory multiple myeloma and del(17p) and/or t(4;14). KW - translocation KW - plus dexamethasone KW - deletion 17P KW - bortezomib KW - therapy KW - abnormalities KW - stem-cell transplantation KW - growth-factor receptor 3 KW - high-risk cytogenetics KW - intergroupe francophone Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-140349 VL - 100 IS - 10 SP - 1327 EP - 1333 ER - TY - JOUR A1 - Terpos, Evangelos A1 - Kleber, Martina A1 - Engelhardt, Monika A1 - Zweegman, Sonja A1 - Gay, Francesca A1 - Kastritis, Efstathios A1 - van de Donk, Niels W. C. J. A1 - Bruno, Benedetto A1 - Sezer, Orhan A1 - Broijl, Annemiek A1 - Bringhen, Sara A1 - Beksac, Meral A1 - Larocca, Alessandra A1 - Hajek, Roman A1 - Musto, Pellegrino A1 - Johnsen, Hans Erik A1 - Morabito, Fortunato A1 - Ludwig, Heinz A1 - Cavo, Michele A1 - Einsele, Hermann A1 - Sonneveld, Pieter A1 - Dimopoulos, Meletios A. A1 - Palumbo, Antonio T1 - European Myeloma Network Guidelines for the Management of Multiple Myeloma-related Complications JF - Haematologica N2 - The European Myeloma Network provides recommendations for the management of the most common complications of multiple myeloma. Whole body low-dose computed tomography is more sensitive than conventional radiography in depicting osteolytic disease and thus we recommend it as the novel standard for the detection of lytic lesions in myeloma (grade 1A). Myeloma patients with adequate renal function and bone disease at diagnosis should be treated with zoledronic acid or pamidronate (grade 1A). Symptomatic patients without lytic lesions on conventional radiography can be treated with zoledronic acid (grade 1B), but its advantage is not clear for patients with no bone involvement on computed tomography or magnetic resonance imaging. In asymptomatic myeloma, bisphosphonates are not recommended (grade 1A). Zoledronic acid should be given continuously, but it is not clear if patients who achieve at least a very good partial response benefit from its continuous use (grade 1B). Treatment with erythropoietic-stimulating agents may be initiated in patients with persistent symptomatic anemia (hemoglobin < 10g/dL) in whom other causes of anemia have been excluded (grade 1B). Erythropoietic agents should be stopped after 6-8 weeks if no adequate hemoglobin response is achieved. For renal impairment, bortezomib-based regimens are the current standard of care (grade 1A). For the management of treatment-induced peripheral neuropathy, drug modification is needed (grade 1C). Vaccination against influenza is recommended; vaccination against streptococcus pneumonia and hemophilus influenza is appropriate, but efficacy is not guaranteed due to suboptimal immune response (grade 1C). Prophylactic aciclovir (or valacyclovir) is recommended for patients receiving proteasome inhibitors, autologous or allogeneic transplantation (grade 1A). KW - bone-disease KW - stem-cell transplantation KW - acute kidney injury KW - erythropoiesis-stimulating agents KW - recombinant-human-erythropoietin KW - randomized controlled trial KW - group consensus statement KW - newly-diagnosed myeloma KW - zoledonic acid KW - enal impairment Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-141913 VL - 100 IS - 10 SP - 1254 EP - 1266 ER - TY - JOUR A1 - Hefner, Jochen A1 - Csef, Herbert A1 - Frantz, Stefan A1 - Glatter, Nina A1 - Warrings, Bodo T1 - Recurrent Tako-Tsubo cardiomyopathy (TTC) in a pre-menopausal woman: late sequelae of a traumatic event? JF - BMC Cardiovascular Disorders N2 - Background “Tako-Tsubo cardiomyopathy” (TTC) is a syndrome characterized by left ventricular (LV) wall motion abnormalities, usually without coronary artery disease, mimicking the diagnosis of acute coronary syndrome. It most often affects post-menopausal women and TTC tends to run a benign course with very low rates of recurrence, complications or mortality. The condition is also called “stress-induced cardiomyopathy” because acute physical or emotional stress appears to be frequently related to its onset. The pathogenic role of premorbid or comorbid psychiatric illnesses has been discussed controversially. For the first time, we present a case of fourfold recurrent TTC with severe complications in a pre-menopausal woman. Furthermore, a long history of flaring posttraumatic stress symptoms anteceded the first event. Case presentation A 43-year old, pre-menopausal Caucasian woman was hospitalized with symptoms of acute coronary syndrome. Clinical examination revealed hypokinetic wall motion in the apical ventricular region with no signs of coronary artery disease and diagnosis of TTC was established. She experienced recurrence three times within the following ten months, which led to thrombembolism and myocardial scarring among others. The circumstances of chronic distress were striking. 16 years ago she miscarried after having removed a myoma according to her doctor’s suggestion. Since then, she has suffered from symptoms of posttraumatic distress which peaked annually at the day of abortion. Chronic distress became even more pronounced after the premature birth of a daughter some years later. The first event of TTC occurred after a family dispute about parenting. Conclusion This is the first case report of fourfold TTC in a pre-menopausal woman. From somatic perspectives, the course of the disease with recurrences and complications underlines the fact that TTC is not entirely benign. Furthermore, it is the first case report of long lasting symptoms of traumatic stress anteceding TTC. Close connections between adrenergic signaling and late onset of clinical stress symptoms are well known in the psychopathology of traumatization. Although larger clinical trials are needed to elucidate possible interactions of premorbid psychiatric illnesses and TTC, cardiologists should be vigilant especially in cases of recurrent TTC. KW - recurrent Tako-Tsubo cardiomyopathy KW - chronic distress KW - gene-environment interaction KW - comprehensive psychosomatic assessment Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-124949 VL - 15 IS - 3 ER - TY - JOUR A1 - Danhof, Sophia A1 - Schreder, Martin A1 - Strifler, Susanne A1 - Einsele, Hermann A1 - Knop, Stefan T1 - Long-Term Disease Control by Pomalidomide-/Dexamethasone-Based Therapy in a Patient with Advanced Multiple Myeloma: A Case Report and Review of the Literature JF - Case Reports in Oncology N2 - Background: Therapy for multiple myeloma (MM) has substantially improved in the era of immunomodulatory drugs and bortezomib. However, the prognosis of patients with progressive disease despite treatment with these ‘novel agents' remains poor. Recently, pomalidomide was approved in this setting, but a median progression-free survival of <4 months still leaves room for improvement. Pomalidomide-based combination therapies are currently under investigation, but data on long-term treatment are lacking. Case Report: We present the case of a 68-year-old woman with refractory MM who received pomalidomide in combination with various drugs including anthracyclines, alkylators and proteasome inhibitors. Initially, major hematological toxicities and infectious complications including a hepatitis B virus reactivation were encountered. With careful dose adjustments and selection of combination partners, pomalidomide treatment was maintained for over 4 years and led to a sustained partial remission. In particular, the well-tolerated regimen of bortezomib, cyclophosphamide and dexamethasone together with pomalidomide was administered for >30 cycles. Conclusion: This case illustrates the value of an individualized approach to myeloma care given an increasing availability of ‘novel agents'. Tailored treatment using these drugs as a backbone is essential to achieve long-lasting responses and minimize side effects. KW - Hepatitis B virus reactivation KW - pomalidomide KW - combination therapy KW - multiple myeloma Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-126093 VL - 8 IS - 1 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 - Mahmood, Zafar A1 - Muhammad, Khalid A1 - Schmalzing, Marc A1 - Roll, Petra A1 - Dörner, Thomas A1 - Tony, Hans-Peter T1 - CD27-IgD- memory B cells are modulated by in vivo interleukin-6 receptor (IL-6R) blockade in rheumatoid arthritis JF - Arthritis Research & Therapy N2 - Introduction Enhanced B cell activity, particularly memory B cells have gained interest in evaluating response during therapies with biologics. CD27-IgD- double-negative (DN) B cells lacking the conventional memory marker CD27 are reported to be part of the memory compartment, however, only scarce data is available for rheumatoid arthritis (RA). We therefore focused on DN B cells in RA, studied their isotypes and modulation during interleukin-6 receptor (IL-6R) inhibition by tocilizumab (TCZ). Methods DN B cells were phenotypically analyzed from 40 RA patients during TCZ at baseline week 12, week 24 and 1 year. A single B cell polymerase chain reaction (PCR) approach was used to study Ig receptors, VH gene rearrangements and specific isotypes. Results Phenotypic analysis showed a significantly expanded population of DN B cells in RA which contain a heterogeneous mixture of IgG-, IgA- and IgM-expressing cells with a clear dominance of IgG+ cells. DN B cells carry rearranged heavy chain gene sequences with a diversified mutational pattern consistent with memory B cells. In contrast to tumor necrosis factor alpha (TNF-α) inhibition, a significant reduction in mutational frequency of BCR gene rearrangements at week 12, 24 and 1 year (P <0.0001) was observed by in vivo IL-6R inhibition. These changes were observed for all BCR isotypes IgG, IgA and IgM at week 12, 24 and 1 year (P <0.0001). IgA-RF, IgA serum level and IgA+ DN B cells decreased significantly (P <0.05) at week 12 and week 24 during TCZ. Patients with a good European League Against Rheumatism (EULAR) response to TCZ had less DN B cells at baseline as compared to moderate responders (P = 0.006). Univariate logistic regression analysis revealed that the frequency of DN B cells at baseline is inversely correlated to a subsequent good EULAR response (P = 0.024) with an odds ratio of 1.48 (95% confidence interval as 1.05 to 2.06). Conclusions In RA, the heterogeneous DN B cell compartment is expanded and dominated by IgG isotype. TCZ can modulate the mutational status of DN Ig isotype receptors over 1 year. Interestingly, the frequency of DN B cells in RA may serve as a baseline predictor of subsequent EULAR response to TCZ. Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-126506 VL - 17 IS - 61 ER - TY - THES A1 - Fließer, Mirjam T1 - Hypoxia and hypoxia-inducible factor 1α modulate the immune response of human dendritic cells against Aspergillus fumigatus T1 - Hypoxie und Hypoxie-induzierbarer Faktor 1α modulieren die Immunantwort humaner dendritischer Zellen gegenüber Aspergillus fumigatus N2 - The mold Aspergillus fumigatus causes life-threatening infections in immunocompromised patients. Over the past decade new findings in research have improved our understanding of A. fumigatus-host interactions. One of them was the detection of localized areas of tissue hypoxia in the lungs of mice infected with A. fumigatus. The transcription factor hypoxia-inducible factor 1α (HIF 1α) is known as the central regulator of cellular responses to hypoxia. Under normoxia, this constitutively expressed protein is degraded by oxygen-dependent mechanisms in most mammalian cell types. Interaction with pathogens can induce HIF 1α stabilization under normoxic conditions in innate immune cells. Bacterial infection models revealed that hypoxic microenvironments and signaling via HIF 1α modulate functions of host immune cells. Moreover, it was recently described that in murine phagocytes, HIF 1α expression is essential to overcome an A. fumigatus infection. However, the influence of hypoxia and the role of HIF 1α signaling for anti-A. fumigatus immunity is still poorly understood, especially regarding dendritic cells (DCs), which are important regulators of anti-fungal immunity. In this study, the functional relevance of hypoxia and HIF 1α signaling in the response of human DCs against A. fumigatus has been investigated. Hypoxia attenuated the pro-inflammatory response of DCs against A. fumigatus during the initial infection as shown by genome-wide microarray expression analyses and cytokine quantification. The up-regulation of maturation-associated molecules on DCs stimulated with A. fumigatus under hypoxia was reduced; however, these DCs possessed an enhanced capacity to stimulate T cells. This study thereby revealed divergent influence of hypoxia on anti-A. fumigatus DC functions that included both, inhibiting and enhancing effects. HIF-1α was stabilized in DCs following stimulation with A. fumigatus under normoxic and hypoxic conditions. This stabilization was partially dependent on Dectin-1, the major receptor for A. fumigatus on human DCs. Using siRNA-based HIF 1α silencing combined with gene expression microarrays, a modulatory effect of HIF-1α on the anti-fungal immune response of human DCs was identified. Specifically, the transcriptomes of HIF-1α silenced DCs indicated that HIF-1α enhanced DC metabolism and cytokine release in response to A. fumigatus under normoxic and hypoxic conditions. This was confirmed by further down-stream analyses that included quantification of glycolytic activity and cytokine profiling of DCs. By that, this study demonstrated functional relevance of HIF 1α expression in DCs responding to A. fumigatus. The data give novel insight into the cellular functions of HIF 1α in human DCs that include regulation of the anti-fungal immune response under normoxia and hypoxia. The comprehensive transcriptome datasets in combination with the down-stream protein analyses from this study will promote further investigations to further characterize the complex interplay between hypoxia, activation of Dectin-1 and HIF-1α signaling in host responses against A. fumigatus. N2 - Der Schimmelpilz Aspergillus fumigatus verursacht lebensbedrohliche Infektionen in immunsupprimierten Patienten. Im letzten Jahrzehnt haben neue Forschungsergebnisse unser Verständnis der Interaktion von A. fumigatus mit seinem Wirt verbessert. Dazu zählt die Beschreibung von lokalisierten Arealen der Hypoxie im Lungengewebe von Mäusen die mit A. fumigatus infiziert wurden. Der Transkriptionsfaktor Hypoxie-induzierbarer Faktor 1α (HIF 1α) ist schon lange als der zentrale Regulator der zellulären Antwort gegenüber Hypoxie bekannt. Unter Normoxie wird dieses konstitutiv exprimierte Protein in den meisten Körperzellen durch sauerstoffabhängige Prozesse abgebaut. In angeborenen Immunzellen kann die Interaktion mit Pathogenen zu einer Stabilisierung von HIF 1α unter normoxischen Bedingungen führen. Bakterielle Infektionsmodelle haben gezeigt, dass hypoxische Mikromilieus und der HIF 1α Signalweg die Funktion von Immunzellen des Wirtes beeinflussen können. Zudem konnte kürzlich nachgewiesen werden, dass die Expression von HIF 1α in murinen Phagozyten während einer Infektion mit A. fumigtus essentiell für eine effektive Bekämpfung des Pilzes ist. Der Einfluss der Hypoxie und die Rolle von HIF 1α für die gegen A. fumigatus gerichtete Immunantwort sind jedoch immer noch unzureichend charakterisiert. Das trifft besonders auf die für die Regulation der anti-fungalen Immunantwort wichtigen dendritischen Zellen (DCs) zu. In dieser Studie wurde die funktionale Bedeutung der Hypoxie und des HIF 1α Signalweges für die Antwort humaner DCs gegenüber A. fumigatus untersucht. Hypoxie hatte einen abschwächenden Effekt auf die initiale pro-inflammatorische Antwort von DCs gegen A. fumigatus. Dies konnte durch genomweite Microarray Expressionsanalysen sowie Zytokinbestimmungen gezeigt werden. Die Hochregulation von Markern, die mit einer Maturierung von mit A. fumigatus-stimulierten DCs assoziiert sind, war unter Hypoxie reduziert. Jedoch zeigten diese DCs eine erhöhte Fähigkeit zur Stimulation von T Zellen. Damit wurden in dieser Studie divergente Effekte der Hypoxie auf die gegen A. fumigatus gerichtete Immunantwort humaner DCs aufgedeckt. Dies beinhaltete sowohl einen inhibierenden als auch einen verstärkenden Einfluss in Abhängigkeit der untersuchten DC Funktion. HIF 1α wurde in DCs nach Stimulation mit A. fumigatus unter normoxischen als auch hypoxischen Bedingungen stabilisiert. Diese Stabilisierung war teilweise abhängig von Dectin-1, dem wichtigsten Rezeptor für A. fumigatus auf humanen DCs. Durch eine Kombination aus RNAi-vermittelter Herunterregulation von HIF 1α und Genexpressions-Microarrays wurde ein modulierender Effekt von HIF 1α auf die anti-fungale Immunantwort humaner DCs identifiziert. Die Transkriptomanalyse von HIF 1α herunterregulierten DCs deutete darauf hin, dass HIF 1α den Metabolismus und die Zytokinfreisetzung in DCs während der Antwort auf A. fumigatus unter normoxischen als auch hypoxischen Bedingungen verstärkt. Dieser Befund wurde durch weiterführende Analysen bestätigt, die eine Quantifizierung der glykolytischen Aktivität sowie die Erstellung eines Zytokinprofils der DCs beinhalteten. Damit konnte in dieser Studie eine funktionale Relevanz der Expression von HIF 1α in DCs für die gegen A. fumigatus gerichtete Immunantwort aufgedeckt werden. Diese Daten geben einen neuen Einblick in die zellulären Funktionen von HIF 1α in humanen DCs, die eine Regulierung der anti-fungalen Immunantwort beinhalten. Die umfassenden Transkriptom-Datensätze dieser Studie, die durch Proteinanalysen funktional ergänzt wurden, bilden die Grundlage für weiterführende Untersuchungen. Damit wird es möglich sein, das komplexe Zusammenspiel aus Hypoxie, Aktivierung von Dectin-1 und Signalübertragung über HIF 1α in der Immunantwort gegen A. fumigatus über die Ergebnisse dieser Studie hinaus noch besser zu charakterisieren. KW - Immunologie KW - Hypoxie KW - Dendritische Zelle KW - Aspergillus fumigatus KW - HIF-1α Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-121392 ER -