TY - THES A1 - Feuchtenberger, Martin T1 - Etablierung einer Fluoreszenz-Multiplex-Polymerase-Ketten-Reaktion zur semiquantitativen und qualitativen Analyse des V H-Repertoires in humanen B-Lymphozyten T1 - Semiquantitative and qualitative assessment of VH-repertoire in human b-lymphocytes by FMPCR N2 - Mit der VH-Fluoreszenz-Multiplex-PCR steht ein neuartiges Screeningverfahren zur semiquantitativen und qualitativen Analyse der VH-Familienverteilung der schweren Kette des Immunglobulinmoleküls in humanen B-Lymphozyten zur Verfügung. In zwei Multiplex-PCRs werden IgH Produkte der sechs häufigsten VH-Familien mit familienspezifischen und fluoreszenzmarkierten Primern (FAM, HEX, NED und ROX) amplifiziert. Die Auftrennung der Produkte erfolgt durch Elektrophorese in einem ABI Prism 377 Sequencer. In verschiedenen Experimenten konnte die hohe Spezifität und Sensitivität der Methode, sowie die gute Reproduzierbarkeit, Reliabilität und Validität der Daten gezeigt werden. Die Analyse des VH-Repertoires gesunder Individuen (n=10) zeigte eine vergleichbare und zugleich charakteristische Häufigkeitsverteilung der VH-Familien unter den Probanden, wie sie auch schon in anderen Studien beschrieben wurde. Die gewonnen Daten wurden ferner durch Sequenzierung von Produkten im Anschluss an eine Single-Cell PCR validiert. Mittels FMPCR konnte darüber hinaus die Existenz monoklonaler B-Lymphozyten bei Patienten mit CLL nachgewiesen werden. Erste Befunde bei Patienten mit Autoimmunerkrankungen zeigten in Einzelfällen ein stark restringiertes B-Zellrepertoire. Die familienspezifische Markierung der Primer mit Fluoreszenzfarbstoffen erlaubt folglich neben der Abschätzung der Häufigkeitsverteilung der VH-Familien auch eine Aussage über die klonale Zusammensetzung des peripheren B-Lymphozytenpools sowie im Falle eines klonalen Peaks die unmittelbare Zuordnung zu einer der sechs VH-Familien. Die FMPCR stellt ein schnell und einfach durchzuführendes, aber zugleich reliables Screeningverfahren zur Analyse des humanen VH-Repertoires bei Erkrankungen mit einer dominierenden Rolle von B-Lymphozyten dar. Dadurch wird es erstmals möglich sein, größere Kollektive von Patienten mit Autoimmunerkrankungen bezüglich eines krankheitsspezifischen Repertoires zu untersuchen. N2 - We established a new tool to perform semiquantitative and qualitative screening for V(H) gene usage frequency during IgH rearrangements in human B-lymphocytes. In two separate multiplex PCRs, the rearranged VDJ regions were amplified with V(H) family-specific primers labeled with different fluorescent dyes (FAM, HEX, NED, or ROX). The relative amount of each of the particular V(H) family products and their ratios were determined by fragment analysis on a ABI PRISM 377 sequencer. We verified that the fluorescent multiplex PCR (FMPCR) shows high specificity and sensitivity, acceptable reproducibility and reliability. Data obtained were well in agreement with results revealed by sequencing following single-cell PCR. Ten healthy volunteers showed a comparable semiquantitative V(H) family distribution. The FMPCR also correctly detected a monoclonal peak in a CLL patient. Thus, labeling primers with various fluorescent dyes allows for an assessment of V(H) family usage and an immediate determination of the involved V(H) gene family if any clonal peaks are present. This method provides a quick, easy, and reliable tool for V(H) repertoire screening of larger populations of patients suffering from diseases with changes in the V(H) repertoire allowing for selection of cases worth a more detailed and cumbersome sequence analysis later on. KW - B-Lymphozyt KW - FMPCR KW - VH-Repertoire KW - Autoimmunität KW - B-lymphocyte KW - FMPCR KW - VH-repertoire KW - autoimmunity Y1 - 2003 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-8426 ER - TY - JOUR A1 - Bittner, Stefan A1 - Bobak, Nicole A1 - Feuchtenberger, Martin A1 - Herrmann, Alexander M A1 - Göbel, Kerstin A1 - Kinne, Raimund W A1 - Hansen, Anker J A1 - Budde, Thomas A1 - Kleinschnitz, Christoph A1 - Frey, Oliver A1 - Tony, Hans-Peter A1 - Wiendl, Heinz A1 - Meuth, Sven G T1 - Expression of K\(_2\)\(_P\)5.1 potassium channels on CD4\(^+\)T lymphocytes correlates with disease activity in rheumatoid arthritis patients JF - Arthritis Research & Therapy N2 - Introduction CD4+ T cells express K2P5.1 (TWIK-related acid-sensitive potassium channel 2 (TASK2); KCNK5), a member of the two-pore domain potassium channel family, which has been shown to influence T cell effector functions. Recently, it was shown that K2P5.1 is upregulated upon (autoimmune) T cell stimulation. The aim of this study was to correlate expression levels of K2P5.1 on T cells from patients with rheumatoid arthritis (RA) to disease activity in these patients. Methods Expression levels of K2P5.1 were measured by RT-PCR in the peripheral blood of 58 patients with RA and correlated with disease activity parameters (C-reactive protein levels, erythrocyte sedimentation rates, disease activity score (DAS28) scores). Twenty patients undergoing therapy change were followed-up for six months. Additionally, synovial fluid and synovial biopsies were investigated for T lymphocytes expressing K2P5.1. Results K2P5.1 expression levels in CD4+ T cells show a strong correlation to DAS28 scores in RA patients. Similar correlations were found for serological inflammatory parameters (erythrocyte sedimentation rate, C-reactive protein). In addition, K2P5.1 expression levels of synovial fluid-derived T cells are higher compared to peripheral blood T cells. Prospective data in individual patients show a parallel behaviour of K2P5.1 expression to disease activity parameters during a longitudinal follow-up for six months. Conclusions Disease activity in RA patients correlates strongly with K2P5.1 expression levels in CD4+ T lymphocytes in the peripheral blood in cross-sectional as well as in longitudinal observations. Further studies are needed to investigate the exact pathophysiological mechanisms and to evaluate the possible use of K2P5.1 as a potential biomarker for disease activity and differential diagnosis. KW - neurology Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-139334 VL - 13 IS - R21 ER - TY - JOUR A1 - Kleinert, Stefan A1 - Roll, Petra A1 - Baumgaertner, Christian A1 - Himsel, Andrea A1 - Burkhardt, Harald A1 - Mueller, Adelheid A1 - Fleck, Martin A1 - Feuchtenberger, Martin A1 - Janett, Manfred A1 - Tony, Hans-Peter T1 - Renal Perfusion in Scleroderma Patients Assessed by Microbubble-Based Contrast-Enhanced Ultrasound N2 - Abstract: Objectives: Renal damage is common in scleroderma. It can occur acutely or chronically. Renal reserve might already be impaired before it can be detected by laboratory findings. Microbubble-based contrast-enhanced ultrasound has been demonstrated to improve blood perfusion imaging in organs. Therefore, we conducted a study to assess renal perfusion in scleroderma patients utilizing this novel technique. Materials and Methodology: Microbubble-based contrast agent was infused and destroyed by using high mechanical index by Siemens Sequoia (curved array, 4.5 MHz). Replenishment was recorded for 8 seconds. Regions of interests (ROI) were analyzed in renal parenchyma, interlobular artery and renal pyramid with quantitative contrast software (CUSQ 1.4, Siemens Acuson, Mountain View, California). Time to maximal Enhancement (TmE), maximal enhancement (mE) and maximal enhancement relative to maximal enhancement of the interlobular artery (mE%A) were calculated for different ROIs. Results: There was a linear correlation between the time to maximal enhancement in the parenchyma and the glomerular filtration rate. However, the other parameters did not reveal significant differences between scleroderma patients and healthy controls. Conclusion: Renal perfusion of scleroderma patients including the glomerular filtration rate can be assessed using microbubble-based contrast media. KW - Medizin KW - Scleroderma KW - renal perfusion KW - contrast-enhanced ultrasound. Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-75207 ER - TY - JOUR A1 - Kleinert, Stefan A1 - Tony, Hans-Peter A1 - Krause, Andreas A1 - Feuchtenberger, Martin A1 - Wassenberg, Siegfried A1 - Richter, Constanze A1 - Röther, Ekkehard A1 - Spieler, Wolfgang A1 - Gnann, Holger A1 - Wittig, Bianca M. T1 - Impact of patient and disease characteristics on therapeutic success during adalimumab treatment of patients with rheumatoid arthritis: data from a German JF - Rheumatology International N2 - The objective of this study was to use data from a noninterventional study to evaluate the effectiveness of adalimumab in rheumatoid arthritis (RA) patients during routine clinical practice and to explore the potential impact of patient and disease characteristics in response to adalimumab therapy. A total of 2,625 RA patients with specified data at baseline (prior to initiating adalimumab treatment) and 12 months entered this study between April 2003 and March 2009. We evaluated response to adalimumab therapy and conducted stepwise regression and subgroup analyses of factors influencing therapeutic response. During the 1-year adalimumab treatment period, disease activity decreased from a baseline mean disease activity score-28 joints (DAS28) of 5.9–3.9, while functional capacity improved from 59.0 to 68.4 Funktionsfragebogen Hannover (FFbH) percentage points. In multivariate regression models, high baseline DAS28 was the strongest positive predictor for decrease in disease activity, and high baseline functional capacity was associated with reduced gains in functional capacity. Male gender was a positive predictor of therapeutic response for both disease activity and functional capacity, while older age and multiple previous biologics were associated with a reduced therapeutic response. Subset analyses provided further support for the impact of baseline DAS28, FFbH, and prior biologic therapy on therapeutic response during treatment. We conclude that treatment with adalimumab leads to decreased disease activity and improved function during routine clinical practice. Patients with high disease activity and low functional capacity are particularly benefitted by adalimumab therapy. KW - antirheumatic agents KW - adalimumab KW - rheumatoid arthritis KW - treatment outcome KW - regression analysis Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-126220 VL - 32 IS - 9 ER - TY - JOUR A1 - Kleinert, Stefan A1 - Tony, Hans-Peter A1 - Krause, Andreas A1 - Feuchtenberger, Martin A1 - Wassenberg, Siegfried A1 - Richter, Constanze A1 - Räther, Ekkehard A1 - Spieler, Wolfgang A1 - Gnann, Holger A1 - Wittig, Bianca M. T1 - Impact of patient and disease characteristics on therapeutic success during adalimumab treatment of patients with rheumatoid arthritis: data from a German noninterventional observational study JF - Rheumatology International N2 - The objective of this study was to use data from a noninterventional study to evaluate the effectiveness of adalimumab in rheumatoid arthritis (RA) patients during routine clinical practice and to explore the potential impact of patient and disease characteristics in response to adalimumab therapy. A total of 2,625 RA patients with specified data at baseline (prior to initiating adalimumab treatment) and 12 months entered this study between April 2003 and March 2009. We evaluated response to adalimumab therapy and conducted stepwise regression and subgroup analyses of factors influencing therapeutic response. During the 1-year adalimumab treatment period, disease activity decreased from a baseline mean disease activity score-28 joints (DAS28) of 5.9–3.9, while functional capacity improved from 59.0 to 68.4 Funktionsfragebogen Hannover (FFbH) percentage points. In multivariate regression models, high baseline DAS28 was the strongest positive predictor for decrease in disease activity, and high baseline functional capacity was associated with reduced gains in functional capacity. Male gender was a positive predictor of therapeutic response for both disease activity and functional capacity, while older age and multiple previous biologics were associated with a reduced therapeutic response. Subset analyses provided further support for the impact of baseline DAS28, FFbH, and prior biologic therapy on therapeutic response during treatment. We conclude that treatment with adalimumab leads to decreased disease activity and improved function during routine clinical practice. Patients with high disease activity and low functional capacity are particularly benefitted by adalimumab therapy. KW - antirheumatic agents KW - arthritis KW - adalimumab KW - rheumatoid KW - treatment outcome KW - regression analysis Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-125118 VL - 32 IS - 9 ER - TY - JOUR A1 - Rech, Juergen A1 - Hueber, Axel J. A1 - Finzel, Stephanie A1 - Englbrecht, Matthias A1 - Haschka, Judith A1 - Manger, Bernhard A1 - Kleyer, Arnd A1 - Reiser, Michaela A1 - Cobra, Jayme Fogagnolo A1 - Figueiredo, Camille A1 - Tony, Hans-Peter A1 - Kleinert, Stefan A1 - Wendler, Joerg A1 - Schuch, Florian A1 - Ronneberger, Monika A1 - Feuchtenberger, Martin A1 - Fleck, Martin A1 - Manger, Karin A1 - Ochs, Wolfgang A1 - Schmitt-Haendle, Matthias A1 - Lorenz, Hanns-Martin A1 - Nuesslein, Hubert A1 - Alten, Rieke A1 - Henes, Joerg A1 - Krueger, Klaus A1 - Schett, Georg T1 - Prediction of disease relapses by multibiomarker disease activity and autoantibody status in patients with rheumatoid arthritis on tapering DMARD treatment JF - Annals of the Rheumatic Diseases N2 - Objective To analyse the role of multibiomarker disease activity (MBDA) score in predicting disease relapses in patients with rheumatoid arthritis (RA) in sustained remission who tapered disease modifying antirheumatic drug (DMARD) therapy in RETRO, a prospective randomised controlled trial. Methods MBDA scores (scale 1-100) were determined based on 12 inflammation markers in baseline serum samples from 94 patients of the RETRO study. MBDA scores were compared between patients relapsing or remaining in remission when tapering DMARDs. Demographic and disease-specific parameters were included in multivariate logistic regression analysis for defining predictors of relapse. Results Moderate-to-high MBDA scores were found in 33% of patients with RA overall. Twice as many patients who relapsed (58%) had moderate/high MBDA compared with patients who remained in remission (21%). Baseline MBDA scores were significantly higher in patients with RA who were relapsing than those remaining in stable remission (N=94; p=0.0001) and those tapering/stopping (N=59; p=0.0001). Multivariate regression analysis identified MBDA scores as independent predictor for relapses in addition to anticitrullinated protein antibody (ACPA) status. Relapse rates were low (13%) in patients who were MBDA-/ACPA-, moderate in patients who were MBDA+/ACPA- (33.3%) and MBDA-ACPA+ (31.8%) and high in patients who were MBDA+/ACPA+ (76.4%). Conclusions MBDA improved the prediction of relapses in patients with RA in stable remission undergoing DMARD tapering. If combined with ACPA testing, MBDA allowed prediction of relapse in more than 80% of the patients. Trial registration number EudraCT 2009-015740-42. KW - Drug-free remission KW - Clinical remission KW - Validation KW - Synovitis Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-187519 VL - 75 IS - 9 ER - TY - JOUR A1 - Stephan, Marlene A1 - Tascilar, Koray A1 - Yalcin-Mutlu, Melek A1 - Hagen, Melanie A1 - Haschka, Judith A1 - Reiser, Michaela A1 - Hartmann, Fabian A1 - Kleyer, Arnd A1 - Hueber, Axel J. A1 - Manger, Bernhard A1 - Figueiredo, Camille A1 - Cobra, Jayme Fogagnolo A1 - Tony, Hans-Peter A1 - Finzel, Stephanie A1 - Kleinert, Stefan A1 - Wendler, Jörg A1 - Schuch, Florian A1 - Ronneberger, Monika A1 - Feuchtenberger, Martin A1 - Fleck, Martin A1 - Manger, Karin A1 - Ochs, Wolfgang A1 - Schmitt-Haendle, Matthias A1 - Lorenz, Hannes Martin A1 - Nüsslein, Hubert A1 - Alten, Rieke A1 - Henes, Joerg A1 - Krüger, Klaus A1 - Schett, Georg A1 - Rech, Jürgen T1 - Physical function of RA patients tapering treatment — a post hoc analysis of the randomized controlled RETRO trial JF - Journal of Clinical Medicine N2 - Several studies have shown that tapering or stopping disease-modifying anti-rheumatic drugs (DMARDs) in rheumatoid arthritis (RA) patients in sustained remission is feasible. However, tapering/stopping bears the risk of decline in physical function as some patients may relapse and face increased disease activity. Here, we analyzed the impact of tapering or stopping DMARD treatment on the physical function of RA patients. The study was a post hoc analysis of physical functional worsening for 282 patients with RA in sustained remission tapering and stopping DMARD treatment in the prospective randomized RETRO study. HAQ and DAS-28 scores were determined in baseline samples of patients continuing DMARD (arm 1), tapering their dose by 50% (arm 2), or stopping after tapering (arm 3). Patients were followed over 1 year, and HAQ and DAS-28 scores were evaluated every 3 months. The effect of treatment reduction strategy on functional worsening was assessed in a recurrent-event Cox regression model with a study-group (control, taper, and taper/stop) as the predictor. Two-hundred and eighty-two patients were analyzed. In 58 patients, functional worsening was observed. The incidences suggest a higher probability of functional worsening in patients tapering and/or stopping DMARDs, which is likely due to higher relapse rates in these individuals. At the end of the study, however, functional worsening was similar among the groups. Point estimates and survival curves show that the decline in functionality according to HAQ after tapering or discontinuation of DMARDs in RA patients with stable remission is associated with recurrence, but not with an overall functional decline. KW - HAQ KW - Rheumatoid Arthritis KW - PROM’s KW - DMARD KW - DAS28 Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-319349 SN - 2077-0383 VL - 12 IS - 11 ER -