Analysis of the antigen-specific T cell response in reactive arthritis by flow cytometry

Arthritis Rheum. 2000 Dec;43(12):2834-42. doi: 10.1002/1529-0131(200012)43:12<2834::AID-ANR25>3.0.CO;2-7.


Objective: In reactive arthritis (ReA) a bacteria-specific T cell response to the triggering microbe is detected in synovial fluid (SF), and an impaired Th1 cytokine response has been described. The recent identification of immunodominant bacterial proteins/peptides and new technologies make a more detailed analysis of the immune response possible. The aim of the present study was to use these new techniques to determine the antigen-specific T cell frequency and the cytokine secretion pattern on stimulation with bacteria-derived recombinant proteins in the peripheral blood (PB) and SF from patients with ReA.

Methods: In 3 patients with Chlamydia-induced ReA and 2 patients with Yersinia-induced ReA, the SF T cell response was investigated after stimulation with the Chlamydia-derived proteins major outer membrane protein (MOMP) and heat-shock protein 60 (Hsp60) and the Yersinia-derived proteins 19-kd protein and Hsp60. In 3 of these patients, the PB T cell response was investigated in parallel. T cells were stimulated in whole blood or whole SF with antigen plus anti-CD28 for 6 hours, brefeldin A was added after 2 hours, and cells were fixed and stained with antibodies against the surface markers CD4 and CD69 and against the cytokines interferon-gamma (IFNgamma), tumor necrosis factor alpha, interleukin-10 (IL-10), and IL-4. Positive cells were quantified by flow cytometry.

Results: In the 3 patients with Chlamydia-induced ReA, the antigen-specific T cell frequency (percentage of IFNgamma CD69 double-positive CD4+ T cells) in response to MOMP (mean +/- SD 1.2 +/- 1.38%) and to Hsp60 (1.21 +/- 1.45%) in SF was about the same. In the 2 patients with Yersinia-induced ReA, the mean +/- SD frequency was 0.66 +/- 0.36% in response to the Hsp60 and 03% +/- 0.22 in response to the 19-kd protein. In the 3 patients whose PB was evaluated, the corresponding T cell response was > or =10 times lower. In 2 patients with Chlamydia-induced ReA, antigen-specific IL-10-positive CD4+ T cells were detected in 0.10-0.23% of the CD4+ T cell subpopulation.

Conclusion: The frequency of antigen-specific T cells to Chlamydia- and Yersinia-derived antigens in the SF of ReA patients is between 1:200 and 1:50. Both the chlamydial Hsp60 and MOMP are dominant T cell antigens in Chlamydia-induced ReA. In patients with Chlamydia-induced ReA, we detected antigen-specific IL-10 secretion, which might mediate an inhibition of effective bacterial clearance.

Publication types

  • Comparative Study

MeSH terms

  • Adult
  • Antigens, Bacterial / blood
  • Arthritis, Reactive / blood*
  • Arthritis, Reactive / immunology
  • Arthritis, Reactive / pathology
  • Chlamydia / immunology
  • Cytokines / metabolism
  • Epitopes / analysis
  • Female
  • Flow Cytometry*
  • Humans
  • Lymphocyte Activation / immunology
  • Male
  • Middle Aged
  • Prohibitins
  • Synovial Fluid / chemistry
  • T-Lymphocytes / immunology*
  • Yersinia / immunology


  • Antigens, Bacterial
  • Cytokines
  • Epitopes
  • PHB2 protein, human
  • Prohibitins