- Meeting abstract
- Open Access
Redox Balance Alterations and Hyporesponsiveness of Synovial T Cells in Rheumatoid Arthritis
© Current Science Ltd 2000
- Published: 15 November 1999
- Rheumatoid Arthritis
- Rheumatoid Arthritis Patient
- Synovial Fluid
- Tyrosine Phosphorylation
- Membrane Localization
In rheumatoid arthritis (RA), the functional status of T lymphocytes is incompletely understood. Synovial fluid (SF) T lymphocytes display phenotypic evidence of former activation, but there is hardly any production of T cell derived cytokines in the synovium. Moreover, the in vitro proliferative responsiveness of SF T lymphocytes is decreased compared with that of peripheral blood (PB) T lymphocytes.
Previous studies have revealed reduced intracellular Ca2+ responses and a decreased overall tyrosine phosphorylation pattern in SF T lymphocytes upon TCR/CD3 simulation . Specifically, the tyrosine phosphorylation of the TCR zeta chain, one of the most proximal events in TCR signaling, was clearly diminished in RA SF T lymphocytes . Moreover, the phosphorylation of a 36kDa protein was virtually absent in RA SF T lymphocytes. Here we report that the 36kDa protein is identified as LAT (linker for activation of T cells). In healthy T lymphocytes, LAT is heavily phosphorylated on tyrosine residues by ZAP-70 (zeta-associated protein of 70kDa) upon TCR engagement, which is required for the activation of PLCg1 and the subsequent influx of Ca²+ . Using FACS analysis, we show that the expression of LAT is reduced in both SF and PB T lymphocytes from RA patients compared with T lymphocytes from healthy controls.
LAT is normally a membrane-bound protein due to the presence of a short α -helical structure. Using immuno-fluorescence staining and microscopy, we found that LAT is displaced from the membrane in SF but not PB T lymphocytes from RA patients. The synovium provides an environment of oxidative stress for the SF T lymphocytes, which are severely depleted in their intracellular levels of the antioxidant glutathione (GSH). The replenishment of GSH in SF T lymphocytes by treatment with NAC (N-acetyl-L-cysteine) restores the membrane localization of LAT, and also the phosphorylation of LAT and the expression of IL-2 after TCR stimulation.
Conclusively, it is demonstrated that the hyporesponsiveness of synovial T lymphocytes in RA is associated with the membrane-displacement of LAT. The membrane localization of LAT is sensitive to intracellular GSH alterations . The displacement of LAT fails to bring ZAP-70 in its proximity after TCR stimulation, whereby LAT remains unphosphorylated and the TCR-mediated signaling pathway is abrogated. Hence, these results suggest a role for the membrane-displacement of LAT in the hyporesponsiveness of SF T lymphocytes as a consequence of local oxidative stress.
- Mirza NM, Relias V, Yunis EJ, Pachas WN, Dasgupta JD: Defective signal transduction via T-cell receptor CD3 structure in T cells from rheumatoid arthritis patients. Hum Immunol. 1993, 36: 91-98. 10.1016/0198-8859(93)90111-D.PubMedView ArticleGoogle Scholar
- Maurice MM, Lankester AC, Bezemer AC, et al: Defective TCR-mediated signaling in synovial T cells in rheumatoid arthritis. J Immunol. 1997, 159: 2973-2978.PubMedGoogle Scholar
- Zhang W, Sloan-Lancaster J, Kitchen J, Trible RP, Samelson LE: LAT: the ZAP-70 tyrosine kinase substrate that links T cell receptor to cellular activation. Cell. 1998, 92: 83-92. 10.1016/S0092-8674(00)80901-0.PubMedView ArticleGoogle Scholar
- Maurice MM, Nakamura H, van der Voort EAM, et al: Evidence for the role of an altered redox state in hyporesponsiveness of synovial T cells in rheumatoid arthritis. J Immunol. 1997, 158: 1458-1465.PubMedGoogle Scholar