Bruno Stuhlmüller - Academia.edu (original) (raw)

Papers by Bruno Stuhlmüller

Journal of Clinical Medicine

Rheumatoid arthritis (RA) synovitis is dominated by monocytes/macrophages with inflammatory patte... more Rheumatoid arthritis (RA) synovitis is dominated by monocytes/macrophages with inflammatory patterns resembling microbial stimulation. In search of triggers, we reduced the intestinal microbiome in 20 RA patients (open label study DRKS00014097) by bowel cleansing and 7-day fasting (≤250 kcal/day) and performed immune monitoring and microbiome sequencing. Patients with metabolic syndrome (n = 10) served as a non-inflammatory control group. Scores of disease activity (DAS28/SDAI) declined within a few days and were improved in 19 of 20 RA patients after breaking the fast (median ∆DAS28 = −1.23; ∆SDAI = −43%) or even achieved remission (DAS28 < 2.6/n = 6; SDAI < 3.3/n = 3). Cytometric profiling with 46 different surface markers revealed the most pronounced phenomenon in RA to be an initially increased monocyte turnover, which improved within a few days after microbiota reduction and fasting. Serum levels of IL-6 and zonulin, an indicator of mucosal barrier disruption, decreased s...

Scientific Reports, 2020

Advances in microbiome research suggest involvement in chronic inflammatory diseases such as rheu... more Advances in microbiome research suggest involvement in chronic inflammatory diseases such as rheumatoid arthritis (RA). Searching for initial trigger(s) in RA, we compared transcriptome profiles of highly inflamed RA synovial tissue (RA-ST) and osteoarthritis (OA)-ST with 182 selected reference transcriptomes of defined cell types and their activation by exogenous (microbial) and endogenous inflammatory stimuli. Screening for dominant changes in RA-ST demonstrated activation of monocytes/macrophages with gene-patterns induced by bacterial and fungal triggers. Gene-patterns of activated B- or T-cells in RA-ST reflected a response to activated monocytes/macrophages rather than inducing their activation. In contrast, OA-ST was dominated by gene-patterns of non-activated macrophages and fibroblasts. The difference between RA and OA was more prominent in transcripts of secreted proteins and was confirmed by protein quantification in synovial fluid (SF) and serum. In total, 24 proteins of...

Annals of the Rheumatic Diseases, 2013

EWRR abstracts rheumatoid arthritis (RA). However, the molecular mechanisms of these agents are p... more EWRR abstracts rheumatoid arthritis (RA). However, the molecular mechanisms of these agents are poorly understood. Objective In order to better understand the specifics effects of Adalimumab and Etanercept, we identified and compared the gene expression profiling in peripheral blood mononuclear cells (PBMCs) from responder (R) RA patients treated by methotrexate (MTX)/ adalimumab (ADA) or MTX/etanercept (ETA) before and at 3 months after the beginning of the treatment. Methods Nineteen RA patients were randomised to receive subcutaneously either ADA (40 mg each other week) or ETA (50 mg per week). Thirteen RA patients [average age: 48.6 ± 14.3 years old (yo), MTX: 13.1 ± 6.3 mg/week (w), initial DAS28: 5.5 ± 0.6] received ADA while six RA patients (age: 52.5 ± 16.1 yo, MTX: 18.3 ± 3 mg/w, initial DAS28: 5.7 ± 1.3) were treated by ETA. The drug efficacy was evaluated with the DAS28 after 3 months of treatment according to the EULAR response criteria in order to discriminate R to NR. A blood sample was carried out before the first injection and after 3 months in order to isolate PBMCs and extract total RNA for hybridisation on whole human genome Agilent 4 × 44 k array. A supervised analysis was performed using t-test (GeneSpring GX software) combined with a fold-change 1.5 in order to identify genes whose expression were altered in PBMCs after 3 months from R to each drug. Next, transcripts sets were analysed to identify potential functional pathways (Human Reactome). Gene expression profiles and functional pathways obtained for R to each anti-TNFα were compared. Results Demographic, clinical and biological characteristics of all the patients were comparable before and after treatment administration. From the R (7/13) patients treated with ADA, a combination of 63 transcripts involved in mTORC1-mediated Signalling was identified. Concerning the R (2/6) treated by ETA, a combination of 16 transcripts involved in TNF Signalling, Death Receptor Signalling was observed. Any overlap was found when we compared these transcripts sets. Moreover, any common biological functions and functional pathways were found between R ADA and R ETA. Conclusions The absence of overlap of transcripts sets and the absence of common pathways leading us to consider that molecular mechanisms of ADA and ETA are specific.

Arthritis & Rheumatism, 1999

The p205 autoantigen and interleukin-2 (IL-2) function synergistically to stimulate T lymphocytes... more The p205 autoantigen and interleukin-2 (IL-2) function synergistically to stimulate T lymphocytes from patients with rheumatoid arthritis (RA), and a p205-derived amino acid sequence is identical to an immunoglobulin sequence located within a domain that is reactive with rheumatoid factors (RF). This study was conducted to analyze in detail the T cell immune response against p205 and to investigate whether immunity to p205 may play a role in T cellmediated immunopathology in active RA. Methods. Cibachron blue, protein A-Sepharose, and gel filtration on Sephacryl were used successively to enrich p205 from synovial fluid (SF). T lymphocytes from RA patients were isolated from the peripheral blood (PB), lymph nodes, and SF, and p205 and peptides derived from known sequences were assessed by T cell proliferation assays in the presence of IL-2. Results. P205-specific proliferation of T cells was observed in PB as well as in SF. When p205 was isolated from RA SF, proliferation of RA T cells peaked on day 3. With p205 purified from SF from trauma patients, there was a significant shift of the maximum T cell proliferation to day 8. T cells were of CD4 or CD8 phenotype, and B cells did not proliferate to a significant degree. The T cell response to p205 was always higher for SF mononuclear cells (SFMC) compared with PBMC (P < < < 0.001). In 1 RA patient who underwent repeated leukapheresis, this led to a reproducible decline in p205-specific T cell proliferation to control levels. PB T cells specifically proliferating in response to p205 were detected in 20 of 32 RA patients (63%). Of 26 patients with other inflammatory rheumatic diseases, only 1 showed a minor response to p205, while normal donors did not demonstrate a significant T cell proliferation. A synthetic p205-derived peptide, with an amino acid sequence identical to an immunoglobulin sequence located in the area where RF binds, was reactive with T cells from RA patients. Conclusion. P205 appears to be a major target of autoreactive T cells in RA. P205-specific T cells are primed and more abundant at the site of inflammation. As a T cell target in RA, p205 may well be an antigen involved in the initiation of RF production. Supported by grants from the Deutsche Forschungsgemeinschaft (Bu 445/4-1 and-2, and SFB421, C6) and the Berlin Regional Arthritis Center.

Background: Most biologics for rheumatoid arthritis (RA) target processes involved in monocyte ac... more Background: Most biologics for rheumatoid arthritis (RA) target processes involved in monocyte activation. To determine when, where and how monocytes become involved in pathogenesis of RA, we analysed monocytes from bone marrow, blood and synovial fluid by gene-expression profiling and cytometry, and[for full text, please go to the a.m. URL]

Copyright information:Taken from "Perspectives and limitations of gene ex... more Copyright information:Taken from "Perspectives and limitations of gene expression profiling in rheumatology: new molecular strategies"Arthritis Research & Therapy 2004;6(4):140-146.Published online 4 Jun 2004PMCID:PMC464885.Copyright © 2004 BioMed Central Ltd

Zeitschrift für Rheumatologie, 2018

ZusammenfassungRheumatische Erkrankungen gehören zu den häufigsten chronisch entzündlichen Krankh... more ZusammenfassungRheumatische Erkrankungen gehören zu den häufigsten chronisch entzündlichen Krankheiten. Neben ausgeprägter Schmerzhaftigkeit und progredienter Gelenkzerstörung reduzieren die rheumatoide Arthritis (RA), die Spondyloarthritiden (SpA) und die Psoriasisarthritis (PsA) die Arbeitsfähigkeit, die Lebensqualität und bei unzureichender Behandlung auch die Lebenserwartung. Seit der Einführung der Biologika zur Therapie dieser Erkrankungen hat die Suche nach geeigneten Biomarkern zur Frühdiagnostik und Vorhersage des Therapieerfolgs zunehmend an Bedeutung gewonnen. Das Hauptziel des Verbundes ArthroMark ist, neue Biomarker zu identifizieren und moderne Bildgebungsverfahren einzusetzen mit dem Ziel, die Diagnose, die Verlaufskontrolle und die Stratifizierung von Patienten mit RA, SpA und PsA zu verbessern. Mit der Entwicklung geeigneter Biomarker für diese Erkrankungen trägt dieses Vorhaben zur Gesundheitsforschung im Bereich chronischer Erkrankungen des Bewegungsapparates bei. Durch die Zusammenarbeit verschiedener nationaler Zentren sollen standortspezifische Ressourcen wie Probenbanken und klinische Studien gemeinsam nutzbar gemacht werden und individuelle Schwerpunkte in der Biomarkeranalyse mit einem entsprechenden Mehrwert vernetzt werden. Gemeinsames Datenmanagement und Vereinheitlichung der Datenerhebung sowie bestmögliche Charakterisierung der Patienten durch neue Bildgebungsmethoden sollen die Qualität der Markerprüfung optimieren.AbstractRheumatic diseases are among the most common chronic inflammatory disorders. Besides severe pain and progressive destruction of the joints, rheumatoid arthritis (RA), spondyloarthritides (SpA) and psoriatic arthritis (PsA) impair working ability, reduce quality of life and if treated insufficiently may enhance mortality. With the introduction of biologics to treat these diseases, the demand for biomarkers of early diagnosis and therapeutic stratification has been growing continuously. The main goal of the consortium ArthroMark is to identify new biomarkers and to apply modern imaging technologies for diagnosis, follow-up assessment and stratification of patients with RA, SpA and PsA. With the development of new biomarkers for these diseases, the ArthroMark project contributes to research in chronic diseases of the musculoskeletal system. The cooperation between different national centers will utilize site-specific resources, such as biobanks and clinical studies for sharing and gainful networking of individual core areas in biomarker analysis. Joint data management and harmonization of data assessment as well as best practice characterization of patients with new imaging technologies will optimize quality of marker validation.

Omics, 2017

Background Targeting molecules involved in monocyte activation is an important treatment strategy... more Background Targeting molecules involved in monocyte activation is an important treatment strategy for RA. In this study we aimed to determine monocyte maturation and activation from bone marrow (BM) via blood into synovial fluid (SF) by investigating monocytes transcriptomes and by cytometric profiling of classical (CD14++CD16-), intermediated (CD14++CD16+) and non-classical (CD14+CD16+) monocytes. Materials and methods CD14+ cells from BM and blood of RA and osteoarthritis (OA) patients were profiled with Affymetrix microarrays. A detailed functional analysis was performed with reference transcriptomes of BM precursors, monocyte blood subsets, monocyte activation and egress from BM induced by G-CSF (granulocyte colony-stimulating factor). Cytometric profiling of CD14, CD16, HLA-DR and CD163 expression were used to determine monocyte subsets and to follow their activation and differentiation in BM, blood and SF. Results Transcriptomes of RA-BM monocytes exhibited i) pronounce gene pattern of early myeloid precursors from BM and ii) weak gene pattern of late myeloid precursors from BM. Transcriptomes of RA blood monocytes demonstrated i) pattern of late myeloid precursors from BM and ii) reduced pattern of terminally differentiated CD14+CD16+ monocytes from blood. Cytometric profiling of BM, blood and SF monocytes in RA and OA showed that all three body compartments have their own distribution of monocyte subsets. BM was characterised with classical and intermediate subsets and both subsets showed decreased CD16 expression in RA when compared to OA. As expected, blood was characterised with three subsets, and RA blood showed decreased CD14 and HLA-DR expression on classical monocytes and reduced frequency of non-classical subset. In RA-SF, classical monocytes were absent, intermediate were most dominant and cell-phenotype with low CD16 expression but similar to non-classical monocytes was related to macrophages. Cell frequency of intermediate subset in SF positively correlated with inflammation (ESR; R>0.85) and showed the highest expression of HLA-DR, CD14, CD163. Conclusions Monocyte turnover is increased in RA and characterised with accelerated monocytopoiesis, faster BM egress and migration into inflamed joints. Permanent monocyte activation in the joint and their role in linking innate and adaptive immunity, which is targeted by biologics, emphasises their high diagnostic value and relevance for therapeutic stratification.

Rheumatoid Arthritis, 2009

Zeitschrift für Rheumatologie, 2012

The introduction of biologics has continuously increased the demand for biomarkers for early diag... more The introduction of biologics has continuously increased the demand for biomarkers for early diagnosis and therapeutic stratification. ArthroMark, a research network funded by the Federal Ministry of Education and Research, aims to establish such biomarkers for rheumatoid arthritis and spondyloarthritides. Biobanks and previous work on genotyping, gene expression and autoreactivity profiling build the basis. Bioinformatic networks will help to harmonize the investigations and a clinical study with modern imaging techniques to characterize the functional relevance of the new biomarkers as effectively as possible. To validate the markers for diagnostic application the network aims to expand gradually.

Journal of Molecular Medicine, 2012

Many cytokines are involved in the pathogenesis of autoimmune diseases and are recognized as rele... more Many cytokines are involved in the pathogenesis of autoimmune diseases and are recognized as relevant therapeutic targets to attenuate inflammation, such as tumor necrosis factor (TNF)-α in rheumatoid arthritis (RA) and interferon (IFN)-α/γ in systemic lupus erythematosus (SLE). To relate the transcriptional imprinting of cytokines in a cell type-and disease-specific manner, we generated gene expression profiles from peripheral monocytes of SLE and RA patients and compared them to in vitro-generated signatures induced by TNF-α, IFN-α2a, and IFN-γ. Monocytes from SLE and RA patients revealed disease-specific gene expression profiles. In vitro-generated signatures induced by IFN-α2a and IFN-γ showed similar profiles that only partially overlapped with those induced by TNF-α. Comparisons between disease-specific and in vitrogenerated signatures identified cytokine-regulated genes in SLE and RA with qualitative and quantitative differences. The IFN responses in SLE and RA were found to be regulated in a STAT1-dependent and STAT1-independent manner, respectively. Similarly, genes recognized as TNF-α regulated were clearly distinguishable between RA and SLE patients. While the activity of SLE monocytes was mainly driven by IFN, the activity from RA monocytes showed a dominance of TNF-α that was characterized by STAT1 down-regulation. The responses to specific cytokines were revealed to be disease-dependent and reflected the interplay of cytokines within various inflammatory milieus. This study has demonstrated that monocytes from RA and SLE patients exhibit disease-specific gene expression profiles, which can be molecularly dissected when compared with in vitro-generated cytokine signatures. The results suggest that an assessment of cytokine-response status in monocytes may be helpful for improvement of diagnosis and selection of the best cytokine target for therapeutic intervention.

Arthritis Research & Therapy, 2000

Genetic associations between rheumatoid arthritis and specific HLA class II genes provide clues t... more Genetic associations between rheumatoid arthritis and specific HLA class II genes provide clues to understanding the molecular basis for disease susceptibility. There is a remarkable structural relationship among different rheumatoid arthritis (RA) susceptibility genes, in which each of the associated class II alleles encodes a sequence of key amino acids termed the 'shared epitope.' Mechanistic models to account for the shared epitope association with RA can be interpreted in the context of an HLA-directed pathway for the development of disease. We suggest that altered T cell activation results from recognition of the shared epitope, providing a potential mechanism by which the shared epitope may be involved in the generation or modulation of self-recognition during antigen presentation and processing. We propose that the shared epitope association with RA is not solely based on a specific peptide binding motif and peptide determinant selection but rather is influenced by a strongly biased direct recognition of shared epitope residues by direct T cell contact. 1. Nepom GT: Major histocompatibility complex-directed susceptibility to rheumatoid arthritis.

Annals of the Rheumatic Diseases, 2015

Objectives: To clarify the factors related to renal atrophy after CS therapy in IgG4-RKD. Methods... more Objectives: To clarify the factors related to renal atrophy after CS therapy in IgG4-RKD. Methods: We retrospectively evaluated clinical features including laboratory data, computed tomography (CT) findings before and after CS therapy in 22 patients diagnosed with IgG4-RKD based on the diagnostic criteria for IgG4-RKD [2], all of whom were followed up for more than 20 months. Results: Sixteen patients were men, and six were women (average age 61.5 years). Average follow-up period was 52.4 months (range 20-93). At diagnosis, their serum IgG4 level was 1043±531 mg/dL, and 7 patients showed hypocomplementemia. The average estimated glomerular filtration rate (eGFR) was 67.9 mL/min/1.73m 2. All patients had extra-renal organ involvement. Multiple low-density lesions (LDLs) on contrast-enhanced CT were observed before CS therapy in all patients. All patients were treated with prednisolone (PSL) at an average initial dose of 35.5±8.4 mg/day (range 20-50), and showed disappearance or reduction of LDLs. At least a part of LDLs resulted in partial renal cortical atrophy in 13 patients (Group A), whereas complete recovery with normal contrast enhancement without a residual cortical scar was achieved in 9 patients (Group B). Pre-treatment eGFR in Group A was significantly lower than that in Group B (60.2±23.4 mL/min/1.73m 2 vs 79.0±19.4 mL/min/1.73m 2 , P=0.035), and the percentage of patients with less than 70 mL/min/1.73m 2 of eGFR in Group A was significantly higher than that in Group B (69.2% vs 22.2%, P=0.040). None of the other factors including age, gender, number of involved organs, serum IgG4 and complement levels, initial dose of PSL, renal function after therapy, recurrence rate of renal lesions, prevalence of diabetes mellitus or hypertension, and follow-up period significantly differed between the two groups. Conclusions: The present study suggests that pre-treatment renal insufficiency may relate to renal atrophy after CS therapy in IgG4-RKD. Whether an early initiation of therapy before renal function declines would prevent the development of renal atrophy remains to be clarified through a larger-scale prospective study. References: [1] Stone JH et al. IgG4-related disease.

Background: Selecting effective drugs for individual patients is the current challenge in rheumat... more Background: Selecting effective drugs for individual patients is the current challenge in rheumatoid arthritis (RA). When methotrexate (MTX) as first line drug fails, targeted therapies with different modes of action may be necessary. Thus, MTX response prediction in early RA provides the opportunity[for full text, please go to the a.m. URL]

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This is a letter to the editor according to the publication from Toonen et al. <br&gt... more This is a letter to the editor according to the publication from Toonen et al. PLoS One. 2012;7(3):e33199. doi: 10.1371/journal.pone.0033199. Epub 2012 Mar 21. https://www.ncbi.nlm.nih.gov/pubmed/22457743

Life, 2020

In rheumatoid arthritis (RA), the expression of many pro-destructive/pro-inflammatory proteins de... more In rheumatoid arthritis (RA), the expression of many pro-destructive/pro-inflammatory proteins depends on the transcription factor AP-1. Therefore, our aim was to analyze the presence and functional relevance of mutations in the coding regions of the AP-1 subunits of the fos and jun family in peripheral blood (PB) and synovial membranes (SM) of RA and osteoarthritis patients (OA, disease control), as well as normal controls (NC). Using the non-isotopic RNAse cleavage assay, one known polymorphism (T252C: silent; rs1046117; present in RA, OA, and NC) and three novel germline mutations of the cfos gene were detected: (i) C361G/A367G: Gln121Glu/Ile123Val, denoted as “fos121/123”; present only in one OA sample; (ii) G374A: Arg125Lys, “fos125”; and (iii) C217A/G374A: Leu73Met/Arg125Lys, “fos73/125”, the latter two exclusively present in RA. In addition, three novel somatic cjun mutations (604–606ΔCAG: ΔGln202, “jun202”; C706T: Pro236Ser, “jun236”; G750A: silent) were found exclusively in...