Interleukin 2, 4 and 5 are sequentially produced in mitogen‐stimulated murine spleen cell cultures

Abstract

Lymphokine production was analyzed in murine spleen lymphocytes stimulated with different T cell mitogens. Using in situ hybridization, frequencies of cells and the kinetics of production of interleukin (IL) 2, 4 and 5 were analyzed. The different mitogens varied in their ability to induce the three interleukins. IL 2 was most successfully induced with a high dose of the calcium ionophore A23187 combined with phorbol 12-myristate 13-acetate (PMA). Significant frequencies of cells containing IL 4 or IL 5 mRNA were found among cells stimulated with an anti-CD3 antibody together with PMA, or pokeweed mitogen. The combination of anti-CD3 and PMA induced relatively high frequencies of all three cytokines. The production was sequential with the highest levels of IL 2 mRNA present during the first 24 h, IL 4 mRNA reaching a peak on day 2 and finally IL 5 peaking on day 3. When cells that had been stimulated with mitogens in vitro were restimulated, the lymphokines were produced more rapidly. The order of production was maintained with IL 2 mRNA reaching a maximum already at 3 h of culture, IL 4 mRNA at 8 h and IL 5 mRNA at 24 h.