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Glutathione Augments in Vitro Proliferative Responses of Lymphocytes to Concanavalin A to a Greater Degree in Old than in Young Rats¹

Laboratory of Immunophysiology, Department of Animal Sciences, College of Agriculture, University of Illinois, Urbana, IL 61801

The proliferative response of splenocytes to concanavalin A (Con A) is significantly lower in aged than in young rats. The addition of reduced (GSH) but not oxidized (GSSG) -glutamylcysteinylglycine (glutathione) in vitro totally reverses the defective proliferative response of splenocytes from aged rats. The inhibitor of -glutamylcysteine synthetase, buthionine sulfoximine, abrogates the in vitro proliferation of splenocytes from young rats activated with Con A, and this effect is significantly reversed by addition of GSH. The enhancing effects of GSH on proliferation of Con A-activated splenocytes of both young and old rats are apparent as early as 24 h and are still evident as late as 72 h in culture. Both cysteine and 2-mercaptoethanol are able to mimic the effect of GSH in causing a greater increase in Con A-induced proliferative responses of splenocytes from old rats (eightfold) as compared to young animals (threefold). Although 2-mercaptoethanol increases expression of membrane-bound transferrin receptors on both Con A-activated splenocytes and T lymphocytes, this alone does not account for the greater enhancement of Con A-induced proliferation of splenocytes from old as compared to young rats. These data are consistent with the hypothesis that the glutathione peroxidase-dependent detoxification system is defective in T lymphocytes of aged rats.

KEY WORDS: • glutathione • 2-mercaptoethanol • buthionine sulfoximine • rat T lymphocyte proliferation • aging

¹ This research was supported by grants to Keith W. Kelley from the National Institutes of Health (AG06246), U.S. Department of Agriculture (89-37265-4536), Office of Naval Research (N00014-89-J-1956) and Moorman Mfg. Co. and in part by Biomedical Shared Instrument Grant No. F10-RR02277 from the National Institutes of Health, which provided a flow cytometer to the Cell Science Laboratory, Biotechnology Center, University of Illinois at Urbana-Champaign.

Manuscript received 19 April 1990. Revision accepted 16 July 1990.

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