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Abstract Number: 0781

Phenotypic and Functional Characterisation of Synovial Fluid-derived Fibroblast-like Synoviocytes in Rheumatoid Arthritis

Ditte Køster1, Johanne Hovgaard Egedal1, Malene Hvid1, Martin Roelsgaard Jakobsen1, Ulf Müller-Ladner2, Bent Deleuran1, Tue Wenzel Kragstrup1, Elena Neumann3 and Morten Aagaard Nielsen1, 1Aarhus University, Aarhus, Denmark, 2Department of Rheumatology, Immunology, Osteology and Physical Medicine, Justus Liebig University Gießen, Campus Kerckhoff, Bad Nauheim, Germany, Bad Nauheim, Germany, 3Department of Rheumatology and Clinical Immunology, Campus Kerckhoff, Justus Liebig University Giessen, Bad Nauheim, Germany

Meeting: ACR Convergence 2020

Keywords: Fibroblasts, Synovial, Inflammation, Mouse Models, RA, rheumatoid arthritis

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Session Information

Date: Saturday, November 7, 2020

Title: RA – Etiology & Pathogenesis Poster

Session Type: Poster Session B

Session Time: 9:00AM-11:00AM

Background/Purpose: Fibroblast-like synoviocytes (FLS) are central cellular components in persistent inflammatory joint diseases such as rheumatoid arthritis (RA). Pathological subsets of FLS have been identified from synovial tissue. However, the synovial tissue obtained from arthroplasty procedures is aquired at late disease stages and the cellular yield obtained from synovial tissue biopsies can be low. Challenging the establishment of human RA in vivo and in vitro models. FLS obtained from the synovial fluid (SF-FLS) are proposed as an alternative source of FLS, but a detailed phenotypical and functional characterization of FLS subsets from the synovial fluid has not been performed.

Methods: In the present study, paired peripheral blood mononuclear cells (PBMC) and SF-FLS from patients with RA were obtained (n=7). FLS were isolated from the synovial fluid by a strict trypsinization protocol and their cellular characteristics and functionality were evaluated at passage 4. Monocultures (SF-FLS) and autologous co-cultures (SF-FLS+PBMC) were established from five patients with RA and subsequently evaluated by flow cytometry, Western blotting and multiplex immunoassays. Human cartilage-sponges (n=3) with SF-FLS and without SF-FLS (n=3) were co-implanted subcutaneously in 15 SCID mice, mice with only cell-free human cartilage-sponges were used as controls (n=12). After 45 days, the implants were evaluated using stained sections to determine the SF-FLS invasion score based on perichondrocytic cartilage degradation. Data are expressed as median (25-75 percentile). P-values < 0.05 were considered statistically significant.

Results: The homogeneous subpopulations of FLS, isolated from the synovial fluid, were negative for CD34 and CD45 [98.9%, (97.5-99.7]) and positive for Thy-1 and PDPN [94.6%, (79.9-97.4]). Without stimulation, RA SF-FLS showed high and comparable levels of NFκB related pathway proteins and secreted multiple pro-inflammatory cytokines and chemokines dominated by IL-6 [2648 pg/mL, (1327-6116)] and MCP-1 [2458 pg/mL, (692-8719)]. SF-FLS increased their ICAM-1 and HLA-DR expression after encountering autologous PBMCs (p< 0.01), (p< 0.05). Further, SF-FLS and PBMC interacted synergistically in a co-culture model of RA and significantly increasing the secretion of several cytokines (IL-1β, IL-2, IL-6, (p< 0.01)) and a chemokine (MCP-1, (p< 0.01)). The invasion score of the human SF-FLS in vivo was at primary site, [1.6, (1.3-1.7)] and contralateral implantation site [1.5, (1.1-2.2)]. The invasion score of the human SF-FLS-containing implants both at primary and contralateral site were significantly higher compared with cartilage-sponges evaluated from SF-FLS-free control mice (p< 0001).

Conclusion: This phenotypical and functional characterization of SF-FLS, do not deviate from synovial tissue FLS and lay a foundation for establishing in vivo and in vitro FLS models. These FLS models will be beneficial in our understanding of the role of this cellular subset in arthritis and for characterization of drugs specifically targeting this pathological RA FLS subset.


Disclosure: D. Køster, None; J. Egedal, None; M. Hvid, None; M. Jakobsen, None; U. Müller-Ladner, Biogen, 8; B. Deleuran, None; T. Kragstrup, Pfizer, 8, Eli Lilly, 8, Novartis, 8, UCB, 8, Gilead, 5, Bristol-Myers Squibb, 5, 8, iBiotech ApS, 4; E. Neumann, None; M. Nielsen, None.

To cite this abstract in AMA style:

Køster D, Egedal J, Hvid M, Jakobsen M, Müller-Ladner U, Deleuran B, Kragstrup T, Neumann E, Nielsen M. Phenotypic and Functional Characterisation of Synovial Fluid-derived Fibroblast-like Synoviocytes in Rheumatoid Arthritis [abstract]. Arthritis Rheumatol. 2020; 72 (suppl 10). https://acrabstracts.org/abstract/phenotypic-and-functional-characterisation-of-synovial-fluid-derived-fibroblast-like-synoviocytes-in-rheumatoid-arthritis/. Accessed September 22, 2023.
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