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OBJECTIVE: To analyse the distribution of FOXP3+CD25+CD4+ regulatory T cells (Treg) in peripheral blood, synovial fluid and tissue of patients with rheumatic disease during relapse and after local treatment. METHODS: FOXP3 expression was assessed by flow cytometry, immunohistochemistry, immunofluorescence and real-time polymerase chain reaction (RT-PCR). The functional suppressive capacity of Treg was analysed after co-culture with effector CD4+CD25- T cells through assessment of proliferation and cytokine secretion. RESULTS: It was shown that FOXP3 protein and mRNA expression in synovial fluid T cells was not confined solely to CD25(bright) T cells as seen in blood, but included CD25(intermediate) and even CD25(neg) T cells. Indeed, synovial fluid CD25(high) T cells showed similar suppressive capacity as CD25(bright) T cells, indicating the presence of functional Treg in T cells with lower intensity of CD25. In synovial tissue, FOXP3+ cells were present in low numbers within T-cell infiltrates and decreased further after intra-articular glucocorticosteroid administration, in parallel with the general reduction in inflammation. CONCLUSIONS: Identification of synovial fluid FOXP3+ Treg with varying intensities of CD25 opens up possibilities for thorough characterisation of this important T-cell subset in the inflammatory compartment. However, only scarce synovial membrane expression of FOXP3 was found even in the absence of overt inflammation, suggesting that the synovial membrane is a site that would benefit therapeutically from Treg expansion.

Original publication

DOI

10.1136/ard.2008.100768

Type

Journal article

Journal

Ann Rheum Dis

Publication Date

12/2009

Volume

68

Pages

1908 - 1915

Keywords

Adult, Aged, Arthritis, Arthritis, Rheumatoid, CD4-Positive T-Lymphocytes, Coculture Techniques, Female, Flow Cytometry, Forkhead Transcription Factors, Gene Expression, Glucocorticoids, Humans, Injections, Intra-Articular, Interleukin-2 Receptor alpha Subunit, Interleukin-7 Receptor alpha Subunit, Male, Middle Aged, RNA, Messenger, Reverse Transcriptase Polymerase Chain Reaction, Synovial Fluid, Synovial Membrane, T-Lymphocyte Subsets, T-Lymphocytes, Regulatory