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FRI0369 Uc-Mscs Inhibit the Differentiation and Proliferation of T Follicular Helper Cell in Rheumatoid Arthritis
  1. R. Liu1,
  2. M. Zhou1,
  3. Y. Sun1,
  4. X. Li2,
  5. X. Feng1,
  6. L. Sun1
  1. 1Department of Rheumatology, The Affiliated Drum Tower hospital of Nanjing University Medical School, Nanjing
  2. 2Department of Immunology, College of Basic Medical Science, Dalian Medical University, Dalian, China

Abstract

Objectives The aim of our study was to elucidate whether umbilical cord derived mesenchymal stem cell (UC-MSCs) had immunomodulatory effect on T follicular helper (Tfh) cell under rheumatoid arthritis (RA) background.

Methods PBMCs that isolated from RA patients and healthy contorl (HC) were cocultured with UC-MSCs at a ratio of 100:1, 10:1 and 1:1 respectively through cell-to-cell contact or in a transwell system at a ratio of 1:10. After 3 days' coculture, PBMCs were collected and the frequency of CD4+CXCR5+PD-1+T cell was examined by flow cytometry. Purified naïve T cells isolated from PBMC of RA patients were cocultured with human UC-MSCs for 4 days at a ratio of 1:10 under Tfh cell-polarizing condition. To detect the effect of MSC on Tfh proliferation, CFSE-labeled purified CD4+ T cells isolated from PBMC of RA patients or HC were stimulated with anti-CD3/CD28 and cocultured with UC-MSCs for 5 days. The frequency of CD4+CXCR5+PD-1+T or CD4+CXCR5+PD-1+AnnexinV+T was examined by flow cytometry and the level of IL-21 in the cultured supernatant was measured by ELISA. The mRNA levels of indoleamine 2,3-dioxygenase (IDO), IL-10, PGE2, HGF, TGF-β and HLA-G in UC-MSCs were tested by RT-PCR. IDO activity of was measured by high-performance liquid chromatography (HPLC), and P-STAT-1/3/5 and p-Akt in UC-MSCs were tested by Western blot. IDO inhibitor 1-MT or anti-IL-10 antibody was added into UC-MSCs and Tfh cell differentiation coculture system for 5 days and then the frequency of CD4+CXCR5+PD-1+T was examined by flow cytometry.

Results UC-MSCs were able to suppress the generation of Tfh cell both in RA patients and in HC in vitro, which was dose-dependent and not relied on cell-to cell contact. UC-MSCs suppressed the differentiation and proliferation of Tfh cell that was more prominent in samples from RA patients, but had no effect on Tfh cell apoptosis. The level of IL-21 in UC-MSCs and Tfh cell differentiation coculture system was significantly decreased. Dramatic increases of both IDO mRNA expression and IDO enzymatic activity were detected in UC-MSCs after coculturing with naïve T cells under Tfh cell-polarizing condition. Meanwhile, pSTAT-1/3/5 and pAkt in these UC-MSCs were increased. The addition of the IDO inhibitor 1-MT, but not anti-IL-10 antibody, could reverse the suppressive effect of UC-MSCs on the differentiation of Tfh cell.

Conclusions UC-MSCs suppress Tfh cell differentiation and proliferation in RA patients, which is partially mediated by the secretion of IDO.

Disclosure of Interest None declared

DOI 10.1136/annrheumdis-2014-eular.2202

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