Characterization of cell-derived microparticles in synovial fluid and plasma of patients with rheumatoid arthritis
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Microparticles (MP) are proposed to play a role in the pathogenesis of rheumatoid arthritis (RA). This study aimed to profile cell lineage-specific MP in patients with RA, osteoarthritis (OA), and healthy controls (HC) in synovial fluid and circulation. Patients with RA (n = 40), OA (n = 30) and HC (n = 33) were included. Cell-free synovial fluid (SF) and platelet-poor plasma samples were stained with annexin V APC and antibodies against CD45, CD20, CD14, CD4, CD8, CD66b, and CD61 for multicolor flow cytometry. Mann–Whitney U test/unpaired T test was used to assess intergroup differences among RA and OA SF and clinical, serological phenotypes of RA based on normality distribution; Kruskal–Wallis test with Dunn’s multiple comparisons for comparing plasma MPs among RA, OA, and HC. Correlation between MP proportions and disease parameters was assessed by Spearman’s correlation. The proportion of annexin V+ MP in SF of patients with RA [5 (6.35)] [median (IQR)] was higher compared to OA [1.8 (1.35), p < 0.001] and plasma of patients with RA [3.45 (5.63)] compared to OA [1.85 (1.4)] and HC [0.9 (1.1), p < 0.001]. Leukocyte-derived [0.85 (1.17)], granulocyte-derived [0.4 (2.05)], monocyte-derived [0.4 (0.4)], and T cell-derived MP [CD4+ – 0.1 (0.1); CD8+ − 0.1(0.1)] were higher in RA SF (p < 0.001). Platelet-derived MP (PMP) were the major fraction [1.5 (4.23), p < 0.001] in RA plasma. Leukocyte-derived MP were higher in RA plasma [0.1 (0.2); p < 0.001) than OA and HC. Annexin V+ MP and PMP were higher in the SF of RA with extra-articular manifestations (n = 15), as compared to those without (n = 25) (p = 0.02; p < 0.01, respectively). High SF granulocyte-derived MP were observed in patients with established RA (n = 24), ACPA-positive RA (n = 32) compared to their negative counterparts (p = 0.03; p = 0.02, respectively). Our observations of higher proportions of cell-derived MP in the plasma and synovial fluid of DMARD-naïve RA patients, their clinical and serological phenotypes suggest their role in dynamic cross talk between the joint and systemic circulation, disease pathology, and progression.
KeywordsRheumatoid arthritis Osteoarthritis Microparticles Synovial fluid Plasma Flow cytometry Autoimmunity
VSN, BNRM, CKG, and KV contributed to the conception and design, acquisition of data, analysis, and interpretation of the data and final approval of the version to be published. BNRM and KV drafted the article. VSN critically revised the article for important intellectual content. VSN, BNRM, CKG, and KV agree to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.
The work was supported by Indian Council of Medical Research (ICMR), India (IRIS ID No.: 2012-2279) and JIPMER Intramural Research Fund (JIP/Res/Intra-PhD/01/2014 and JIP/Res/Intra-PhD/02/2015-16).
Compliance with ethical standards
Conflict of interest
Ms. Benita NR Michael, Mr. Vallayyachari Kommoju, Dr. Chengappa Kavadichanda Ganapathy, and Dr. Vir Singh Negi declare that they have no conflict of interest.
The study was approved by the JIPMER institute ethics committee and conducted the following Principles of the Declaration of Helsinki (1964) and its later amendments or comparable ethical standards. Protocol No. JIP/IEC/2013/1/107 dated 15.03.2013.
Informed consent was obtained from all individual participants included in the study.
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