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

Clin Exp Immunol. 2002 Aug;129(2):370-8. doi: 10.1046/j.1365-2249.2002.01868.x.


In this study we examined the cytokine production by T cells and TCRVbeta 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-gamma- 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-gamma, 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-Vbeta families may have the potential to modulate the immune response in RA patients.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Arthritis, Rheumatoid / genetics
  • Arthritis, Rheumatoid / immunology*
  • Case-Control Studies
  • Cytokines / biosynthesis*
  • Cytokines / blood
  • Female
  • Flow Cytometry
  • Gene Rearrangement, beta-Chain T-Cell Antigen Receptor
  • Genes, T-Cell Receptor
  • Humans
  • Interferon-gamma / biosynthesis
  • Interferon-gamma / blood
  • Interleukin-2 / biosynthesis
  • Interleukin-2 / blood
  • Interleukin-4 / biosynthesis
  • Interleukin-4 / blood
  • Interleukin-5 / biosynthesis
  • Interleukin-5 / blood
  • Male
  • Middle Aged
  • Synovial Fluid / cytology
  • Synovial Fluid / immunology
  • T-Lymphocyte Subsets / immunology*


  • Cytokines
  • Interleukin-2
  • Interleukin-5
  • Interleukin-4
  • Interferon-gamma