Simultaneous analysis of T cell clonality and cytokine production in rheumatoid arthritis using three-colour flow cytometry

Bakakos, P., Pickard, C., Wong, W.M., Ayre, K.R., Madden, J., Frew, A.J., Hodges, E., Cawley, M.I. and Smith, J.L. (2002) Simultaneous analysis of T cell clonality and cytokine production in rheumatoid arthritis using three-colour flow cytometry. Clinical and Experimental Immunology, 129 (2), 370-378. (doi:10.1046/j.1365-2249.2002.01868.x).

Abstract

In this study we examined the cytokine production by T cells and TCRV? subsets in peripheral blood (PB) and synovial fluid (SF) from six RA patients and PB from 10 normal subjects, using three-colour flow cytometry. In two RA subjects we assessed T cell clonality by RT PCR using TCRBV family-specific primers and analysed the CDR3 (complementarity determining region 3) length by GeneScan analysis. A high percentage of IFN-?- and IL-2- producing cells was observed among the PB T cells in both the RA patients and normal controls and among the SF T cells in RA patients. In contrast, the percentage of T cells producing IL-4 and IL-5 was small among PB T cells in both RA patients and normal controls and among SF T cells in RA patients.
There was no significant difference in the production of IFN-?, IL-2 and IL-5 between the two compartments (PB and SF); however, there were significantly more IL-4-producing cells in SF. Molecular analysis revealed clonal expansions of four TCRBV families in SF of two of the RA patients studied: TCRBV6·7, TCRBV13·1 and TCRBV22 in one and TCRBV6·7, TCRBV21·3 and TCRBV22 in the second. These expansions demonstrated cytokine expression profiles that differed from total CD3+ cells, implying that T cell subsets bearing various TCR-V? families may have the potential to modulate the immune response in RA patients.

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