Osteoarthritis (OA) is a heterogeneous disease which rheumatologists consider to be noninflammatory. However, recent studies suggest that, at least in certain patients, OA is an inflammatory disease and that patients often exhibit inflammatory infiltrates in the synovial membranes (SMs) of macrophages and activated T cells expressing proinflammatory cytokines. We report here that the expression of CD3zeta is significantly decreased in T cells infiltrating the SMs of patients with OA. The CD3zeta chain is involved in the T-cell signal transduction cascade, which is initiated by the engagement of the T-cell antigen receptor and which culminates in T-cell activation. Double immunofluorescence of single-cell suspensions derived from the SMs from nine patients with OA revealed significantly increased proportions of CD3epsilon-positive (CD3epsilon+) cells compared with the proportions of CD3zeta-positive (CD3zeta+) T cells (means +/- standard errors of the means, 80.48% +/- 3.92% and 69.02% +/- 6.51%, respectively; P = 0.0096), whereas there were no differences in the proportions of these cells in peripheral blood mononuclear cells (PBMCs) from healthy donors (94.73% +/- 1.39% and 93.79% +/- 1.08%, respectively; not significant). The CD3zeta+ cell/CD3epsilon+ cell ratio was also significantly decreased for T cells from the SMs of patients with OA compared with that for T cells from the PBMCs of healthy donors (0.84 +/- 0.17 and 0.99 +/- 0.01, respectively; P = 0.0302). The proportions of CD3epsilon+ CD3zeta+ cells were lower in the SMs of patients with OA than in the PBMCs of healthy donors (65.04% +/- 6.7% and 90.81% +/- 1.99%, respectively; P = 0.0047). Substantial proportions (about 15%) of CD3epsilon+ CD3zeta-negative (CD3zeta-) and CD3epsilon-negative (CD3epsilon-) CD3zeta- cells were found in the SMs of patients with OA. Amplification of the CD3zeta and CD3delta transcripts from the SMs of patients with OA by reverse transcriptase PCR consistently exhibited stronger bands for CD3delta cDNA than for CD3zeta cDNA The CD3zeta/CD3delta transcript ratio in the SMs of patients with OA was significantly lower than that in PBMCs from healthy controls (P < 0.0001). These results were confirmed by competitive MIMIC PCR. Immunoreactivities for the CD3zeta protein were detected in the SMs of 10 of 19 patients with OA, and they were of various intensities, whereas SMs from all patients were CD3epsilon+ (P = 0.0023). The decreased expression of the CD3zeta transcript and protein in T cells from the SMs of patients with OA relative to that of the CD3epsilon transcript is suggestive of chronic T-cell stimulation and supports the concept of T-cell involvement in OA.