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 ([Ca(2+)](i)) is one of the earliest events to occur in T lymphocytes following stimulation of the transmembrane T cell receptor/CD3 complex (TCR/CD3). This [Ca(2+)](i) mobilization has been found to be sensitive to intracellular thiol redox status, which in turn is modulated by cellular glutathione (GSH) content. We have previously reported that GSH depletion, by treatment with either the alpha, beta-carbonyl diethyl maleate or the aromatic halo-compound 1-chloro-2,4-dinitrobenzene, correlates with decreased [Ca(2+)](i) mobilization in anti-CD3 monoclonal antibody (mAb)-stimulated human peripheral blood lymphocytes (HPBL). This prompted us to determine whether this correlation between GSH content and TCR/CD3 signal transduction capability was also present in murine lymphocytes, since the mouse model is often used as a surrogate for the human immune system. The results 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 depleted of GSH by greater than 40% were found to have a decreased [Ca(2+)](i) mobilization following anti-CD3 mAb stimulation. Similar to what has been described for HPBL, these results indicate that the cellular GSH status influences the initial response of murine T lymphocytes to TCR/CD3 stimulation.