Agents which block membrane lipid peroxidation enhance mouse spleen cell immune activities in vitro: relationship to the enhancing activity of 2‐mercaptoethanol

Abstract

A broad spectrum of agents known to block various steps in the lipid peroxidation process were tested for their ability to protect mouse spleen cells and thereby enhance their activities, in vitro, in either the primary antibody response or the lipopolysaccharide-stimulated proliferation response. Each agent (superoxide dismutase, butylated hydroxyanisole/butylated hydroxytoluene/n-propyl gallate, lucigenin, and alpha-tocopherol) was able to enhance the cellular response in both assay systems. The degree of enhancement of these immune functions was in proportion to the efficacy of each agent in blocking the overall process in lipid peroxidation. Previous work in this laboratory has shown that the enhancement of the primary antibody response by 2-mercaptoethanol (2-ME) is mediated by the enhanced availability of reduced glutathione in the culture medium. Suboptimal doses of each lipid antioxidant agent were able to enhance the antibody response in the additive manner with a suboptimal dose of 2-ME up to a maximum response equal to that achieved with an optimal dose of 2-ME alone. These data support the hypothesis that the enhancement of cellular responses in the presence of 2-ME is mediated by the lipid antioxidant activity of reduced glutathione.

Effect of Vitamin E on Contact Sensitization Responses Induced by 2,4-Dinitrochlorobenzene in Mice.

“…2-Mercaptoethanol (2-ME) is usually added to the culture medium for in vitro lymphocyte proliferation assay, as a synthetic antioxidant agent to enhance the (cellular) response (27,28). This enhancement was mediated by the enhanced availability of glutathione to prevent cell membrane lipid peroxidation (29). Oonishi et al found that, in the presence of 2-ME, there was no enhancement of proliferation by a-tocopherol (30).…”

Involvement and relative importance of at least two distinct mechanisms in the effects of 2‐mercaptoethanol on murine lymphocytes in culture

“…Early investigations of the mechanism of action of 2-ME emphasized its antioxidant properties (Toohey, 1975;Hoffeld, 1981). However, the observation that the oxidized (disulfide) form of 2-ME could be as effective as the reduced form (Ohmori and Yamamoto, 1983) indicated that direct antioxidant actions alone could not explain the effect of 2-ME on cultured lymphocytes.…”

Antibody production in cultured blood lymphocytes from breast cancer patients

“…We added transferrin, as suggested by Iscove and Melchers (1978), and reduced GSH, whose high content in foetal calf serum (FCS) accounts for its usefulness in murine cell cultures (Hoffeld & Oppenheim, 1980) and whose promoting activity on human PBL proliferation has been recently demonstrated (Hoffeld, 1981;Noelle & Lawrence, 1981). GSH, together with 2-ME, which prevents its spontaneous oxidation, allowed us to eliminate FCS and avoid the use of a special low oxygen-tension mixture in the incubation chamber.…”