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Alterations in Immune Function in Rats Caused by Dietary Lipotrope Deficiency: Effect of Culture Medium, 2-Mercaptoethanol and Mitogen Dose on the In Vitro Lymphocyte Transformation Response¹

Department of Nutrition and Food Science, Massachusetts Institute of Technology, Cambridge, MA 02139

The effect of variations in culture media, mitogen dose, harvest time, 2-mercaptoethanol (2-ME) addition and length of [³H]thymidine pulse on the concanavalin A (Con A)-stimulated splenic lymphocyte transformation response of male, weanling Sprague-Dawley rats maintained on a control (C), folacin-deficient (F) or marginal methionine-choline (M/C) diet was examined. Spenocytes cultured in minimal essential medium reached an optimal response on day 4. The response of F rats was significantly lower than C rats on day 3 and day 4. The response of M/C rats was lower on day 3. The addition of 2-ME to the culture medium shifted the cell cycle kinetics toward an earlier peak response time (day 2), and significant differences among groups were seen only under suboptimal conditions. Cells cultured in Medium 199 had a low transformation response, which reached peak stimulation on day 5. The response of F and M/C rats was significantly lower than C animals on days 3 and 4. In contrast, splenocytes cultured in medium 199 + 2-ME reached optimal stimulation on day 2, with no significant difference between groups. No effect on cell viability was seen from 2-ME, but it did accelerate cell cycle kinetics and reversed the normal age-induced immunosuppression seen in C animals

KEY WORDS: • immunocompetence • lipotrope • folacin • methionine-choline • 2-mercaptoethanol

¹ This investigation was supported by Public Health Service Grant No. 5R22-HDO6917 awarded by the National Cancer Institute, Department of Health and Human Services.

² Present address: State University of New York Upstate Medical Center, College of Medicine, Syracuse, NY 13210.

Manuscript received 13 April 1982.