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 response.
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. Splenocytes 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 were lower on day 3. The addition of 2-ME to the culture medium shifted the cell cycle kinetics toward and 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 significantly differences 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.
D000818:Animals; D002478:Cells, Cultured; D002796:Choline Deficiency / Q000276:immunology*; D003208:Concanavalin A / Q000494:pharmacology; D003470:Culture Media; D004305:Dose-Response Relationship, Drug; D005494:Folic Acid Deficiency / Q000276:immunology*; D008198:Lymph Nodes / Q000166:cytology; D008213:Lymphocyte Activation / Q000187:drug effects*; D008297:Male; D008623:Mercaptoethanol / Q000494:pharmacology; D008715:Methionine / Q000172:deficiency*; D051381:Rats; D011919:Rats, Inbred Strains; D013154:Spleen / Q000166:cytology; D013936:Thymidine / Q000378:metabolism; D013950:Thymus Gland / Q000166:cytology
D003470:Culture Media; D003208:Concanavalin A; D008623:Mercaptoethanol; D008715:Methionine; D013936:Thymidine