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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 activated cells were confirmed in RA-SF compared to OA-SF and some like CXCL13, CCL18, S100A8/A9, sCD14, LBP reflected this increase even in RA serum. Consequently, pathogen-like response patterns in RA suggest that direct microbial influences exist. This challenges the current concept of autoimmunity and immunosuppressive treatment and advocates new diagnostic and therapeutic strategies that consider microbial persistence as important trigger(s) in the etiopathogenesis of RA.

Zusammenfassung Hintergrund Patient-reported-outcomes (PROs) haben einen zunehmenden Einfluss auf die unmittelbare Therapieentscheidung des Arztes in der klinischen Praxis. Zielstellung Überprüfung der Integration und des Nutzens von elektronisch erhobenen PROs per ScoreCheck Rheuma®-App (SCR App) im klinischen Praxisalltag der rheumatologischen Fachambulanz (Dispensaire) der Charité - Universitätsmedizin Berlin in Hinblick auf Praxistauglich, Patientenakzeptanz und medizinischen Mehrwert. Patienten und Methoden N=190 Patienten (63% weiblich, mittleres Alter: 55 Jahre, min 23-max 82; Diagnosen: 52,9% Rheumatoide Arthritis, 28,6% Spondyloarthritis, 9,3% Kollagenosen, 3,6% Vaskulitis, 5,6% sonstiges) sowie 3 medizinische Fachangestellte (MFA) und 3 Ärzte (Rheumatologen) haben an diesem Pilotprojekt teilgenommen. Die folgenden PROs wurden elektronisch mittels SCR App von den eingeschlossenen Patienten ausgefüllt: FFbH (Funktionsfragebogen Hannover), BASDAI (Bath Ankylosing Spondylitis Disease Activity Index) und BASFI (Bath Ankylosing Spondylitis Functional Index) sowie ein Fragebogen zur Selbsteinschätzung. Zusätzlich wurde für jeden eingeschlossenen Patienten je ein Beurteilungs-Protokoll vom Patienten selbst, von der zuständigen MFA und vom betreuenden Arzt ausgefüllt. Ergebnisse Von den 190 teilnehmenden Patienten gaben 147 (77%) an, Vorerfahrungen mit Touchpad-Geräten zu haben. 96% der Patienten fühlten sich außerdem gut bis sehr gut in die Bedienung des iPADs und der SCR App durch die MFAs eingewiesen und fanden das Ausfüllen der Fragebögen auf dem iPAD einfach; Schwierigkeiten, wie z. B. mit der Schriftgröße, der Handhabung, usw. wurden nur von 10% der Patienten angegeben. Immerhin 74% der Patienten wünschten sich den vermehrten Einsatz elektronischer Hilfsmittel beim Arztbesuch und 75% bevorzugten den Einsatz des iPADs gegenüber dem Ausfüllen der Fragebögen auf Papier. Die MFAs gaben in über 80% der Fälle an, dass die Patienten die Fragebögen zu den PROs ohne oder mit nur wenig Hilfe selbstständig ausfüllen konnten. In ca. 2/3 der Fälle sahen die MFAs Vorteile durch die Anwendung der SCR App sowohl bezüglich der Qualität der Patientenbetreuung als auch bezüglich der Erfassung der Funktionsscores. Die Ärzte sahen in 2/3 der Fälle eher keine Zeitersparnis durch den Einsatz des iPADs. In 78% der Fälle konnte der Patientenscore in das Arzt-Patienten-Gespräch eingebracht werden, jedoch sparte die Anwendung der SCR App nur in 50% der Fälle Zeit im Praxisalltag. Die Ärzte sahen v. a. Vorteile bezüglich der Anamneseerhebung und der Therapieempfehlungen. Schlussfolgerung Insgesamt zeigt das vorgestellte Pilotprojekt zur Anwendung und Integration der SCR App eine gute Resonanz bei den Patienten, MFAs und Ärzten. Die Arztbewertung ergibt, dass durch die SCR App bei jedem zweiten Patienten ein Vorteil bei der Therapieempfehlung gesehen wurde.

ObjectiveRheumatoid arthritis (RA) accompanies infiltration and activation of monocytes in inflamed joints. We investigated dominant alterations of RA monocytes in bone marrow (BM), blood and inflamed joints.MethodsCD14+ cells from BM and peripheral blood (PB) of patients with RA and osteoarthritis (OA) were profiled with GeneChip microarrays. Detailed functional analysis was performed with reference transcriptomes of BM precursors, monocyte blood subsets, monocyte activation and mobilisation. Cytometric profiling determined monocyte subsets of CD14++CD16−, CD14++CD16+ and CD14+CD16+ cells in BM, PB and synovial fluid (SF) and ELISAs quantified the release of activation markers into SF and serum.ResultsInvestigation of genes differentially expressed between RA and OA monocytes with reference transcriptomes revealed gene patterns of early myeloid precursors in RA-BM and late myeloid precursors along with reduced terminal differentiation to CD14+CD16+monocytes in RA-PB. Patterns associated with tumor necrosis factor/lipopolysaccharide (TNF/LPS) stimulation were weak and more pronounced in RA-PB than RA-BM. Cytometric phenotyping of cells in BM, blood and SF disclosed differences related to monocyte subsets and confirmed the reduced frequency of terminally differentiated CD14+CD16+monocytes in RA-PB. Monocyte activation in SF was characterised by the predominance of CD14++CD16++CD163+HLA-DR+ cells and elevated concentrations of sCD14, sCD163 and S100P.ConclusionPatterns of less mature and less differentiated RA-BM and RA-PB monocytes suggest increased turnover with accelerated monocytopoiesis, BM egress and migration into inflamed joints. Predominant activation in the joint indicates the action of local and primary stimuli, which may also promote adaptive immune triggering through monocytes, potentially leading to new diagnostic and therapeutic strategies.

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.

Objectives Proliferative lupus nephritis (LN) is one of the major concerns in the treatment of systemic lupus erythematosus (SLE). Here we evaluate urinary CD4 T cells as a biomarker of active LN and indicator of treatment response. Methods Urinary CD3CD4 T cells were quantified using flow cytometry in 186 urine samples from 147 patients with SLE. Fourteen patients were monitored as follow-up. Thirty-one patients with other nephropathies and 20 healthy volunteers were included as controls. Results In SLE, urinary CD4 T cell counts ≥800/100 ml were observed exclusively in patients with active LN. Receiver operator characteristic analysis documented clear separation of SLE patients with active and non-active LN (area under the curve 0.9969). All patients with up-to-date kidney biopsy results showing proliferative LN had high urinary CD4 T cell numbers. In patients monitored under therapy, normalisation of urinary CD4 T cell counts indicated lower disease activity and better renal function. In contrast, patients with persistence of, or increase in, urinary T cells displayed worse outcomes. Conclusions Urinary CD4 T cells are a highly sensitive and specific marker for detecting proliferative LN in patients with SLE. Furthermore, monitoring urinary CD4 T cells may help to identify treatment responders and treatment failure and enable patient-tailored therapy in the future.

To test the hypothesis that microanatomical differences in joint disease localisation could be exploited using high-resolution MRI to better differentiate among rheumatoid arthritis (RA), spondyloarthritis/psoriatic arthritis (SpA/PsA) and osteoarthritis (OA) in clinical practice. Sixty-nine patients with suspected inflammatory joint disease of the hand or feet underwent high-resolution MRI using a small loop coil. Images were scored blinded to the clinical status. Various joint changes like periostitis, osteitis, erosions, enthesitis and synovitis were recorded. The image-based diagnosis was compared with the clinical diagnosis. In 59.4 % of the patients the clinical diagnosis was confirmed on image analysis. This was high for OA (80 %), moderately good for RA (67 %) but only 50 % for SpA/PsA. The major difficulty was to distinguish OA from SpA/PsA where common imaging findings are evident including periostitis (SpA/PsA 45 %, OA 40 % compared with RA 0 %; P = 0.015). Likewise, osteitis was frequently detected in SpA/PsA (79 %) and OA (80 %) and less frequently in RA (42 %) (P = 0.014). Characterisation of inflammatory disorders of small joints merely using high-resolution MRI remains challenging especially in the differentiation between OA and PsA. These findings are likely explained by common microanatomical similarities in disease expression rather than limitations of imaging techniques. • High-resolution MRI is increasingly used to investigate joint disease. • Osteitis and periostitis occur in psoriatic and osteoarthritis (but not rheumatoid arthritis). • In severely affected patients the amount of synovitis and erosions is similar.