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Limiting dilution analysis of IL-2 production: studies of age, genotype, and regulatory interactions
We have previously described a limiting dilution method for estimating both the frequency of mouse T helper cells that can respond to alloantigens by producing Interleukin-2 (IL-2) and the amount of IL-2 produced by each responding cell (together with its progeny). At these very low cell concentrations, the amount of IL-2 produced per cell is remarkably higher (30-300 X) than the amounts generated under conventional, high density, culture conditions. We show here that cells cultured at intermediate densities "regulate" the amounts of IL-2 produced, so that the IL-2 concentration becomes independent of helper cell dose. The much higher level of IL-2 produced at very low concentrations therefore probably represents the loss, by dilution, of a regulatory cell that is less frequent than the IL-2 producing cells themselves. We show also how limiting dilution methods can be applied to KLH-responsive IL-2 secreting cells, and to questions about the effects of genotype and age on IL-2 production (Key words: Interleukin-2, helper T cells, aging, suppressor cells).
We have previously described a limiting dilution method for estimating both the frequency of mouse T helper cells that can respond to alloantigens by producing Interleukin-2 (IL-2) and the amount of IL-2 produced by each responding cell (together with its progeny). At these very low cell concentrations, the amount of IL-2 produced per cell is remarkably higher (30-300 X) than the amounts generated under conventional, high density, culture conditions. We show here that cells cultured at intermediate densities "regulate" the amounts of IL-2 produced, so that the IL-2 concentration becomes independent of helper cell dose. The much higher level of IL-2 produced at very low concentrations therefore probably represents the loss, by dilution, of a regulatory cell that is less frequent than the IL-2 producing cells themselves. We show also how limiting dilution methods can be applied to KLH-responsive IL-2 secreting cells, and to questions about the effects of genotype and age on IL-2 production (Key words: Interleukin-2, helper T cells, aging, suppressor cells).