Analysis by in situ hybridization of cells expressing mRNA for interleukin 4 in the developing thymus and in peripheral lymphocytes from mice

Abstract

We have made use of RNA.RNA in situ hybridization to study the presence of cells producing mRNA for interleukin 4 (IL-4) in the developing thymus, spleen, and T-cell line 2.19. Approximately 1 of 300-400 spleen cells expressed detectable IL-4 mRNA 24 hr after their stimulation by the lectin concanavalin A. Spleen cells were also induced to express mRNA for IL-4 by stimulation with alloantigens. Splenocytes producing mRNA for IL-4 were detected 4 hr after stimulation by concanavalin A; the response peaked at approximately equal to 24 hr and was undetectable by 72 hr. Cyclosporin A inhibited the synthesis of IL-4 mRNA in the T-cell line 2.19, which had been induced by concanavalin A. Approximately 1 of 10 fetal thymocytes at day 14 of gestation expressed mRNA for IL-4 after their stimulation by phorbol 12-myristate 13-acetate and ionomycin. Both the frequency of fetal thymocytes expressing IL-4 mRNA and the amount of mRNA for IL-4 synthesized per cell sharply decreased at day 16 of gestation, and less than 1 of 1800 fetal thymocytes at day 18 of gestation expressed detectable IL-4 mRNA. Our results define the relative frequency of cells capable of expressing IL-4 mRNA after stimulation in vitro in the spleen and in the developing thymus. The data strongly argue for an important role of IL-4 in growth and differentiation of lymphoid cells, notably during T-cell development within the thymus.