THE EFFECT OF IN-VITRO PHORONE EXPOSURE ON GLUTATHIONE CONTENT AND T-CELL ANTIGEN RECEPTOR (CD3)-STIMULATED CALCIUM MOBILIZATION IN MURINE SPLENIC T-LYMPHOCYTES

An increase in cytosolic free calcium ([Ca2+](i)) is one of the earlie st events to occur in T lymphocytes following stimulation of the trans membrane T cell receptor/CD3 complex (TCR/CD3). This [Ca2+](i) mobiliz ation has been found to be sensitive to intracellular thiol redox stat us, which in turn is modulated by cellular glutathione (GSH) content. We have previously reported that GSH depletion, by treatment with eith er the alpha, beta-farbonyl diethyl maleate or the aromatic halo-compo und 1-chloro-2,4-dinitrobenzene, correlates with decreased [Ca2+](i) m obilization in anti-CD3 monoclonal antibody (mAb)-stimulated human per ipheral blood lymphocytes (HPBL). This prompted us to determine whethe r this correlation between GSH content and TCR/CD3 signal transduction capability was also present in murine lymphocytes, since the mouse mo del is often used as a surrogate for the human immune system. The resu lts presented here demonstrate that in vitro treatment with the alpha, beta-carbonyl phorone dose-dependently depletes intracellular GSH in murine splenic T lymphocytes. Both CD4(+) and CD8(+) T lymphocytes dep leted of GSH by greater than 40% were found to have a decreased [Ca2+] (i) mobilization following anti-CD3 mAb stimulation. Similar to what h as been described for HPBL, these results indicate that the cellular G SH status influences the initial response of murine T lymphocytes to T CR/CD3 stimulation. (C) 1997 Elsevier Science Ltd.