Secretion of IL-4 from human basophils: The relationship between IL-4 mRNA and protein in resting and stimulated basophils

Donald MacGlashan, Jane M. White, Shau Ku Huang, Santa J. Ono, John T. Schroeder, Lawrence M. Lichtenstein

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Abstract

Purified human basophils have been examined for secretion of IL-4 protein and expression of IL-4 mRNA after stimulation with several secretagogues. In general, these studies used a 15-min preincubation with IL-3, before challenge with secretagogues. Under these conditions, IL-4 release averaged 30 pg/106 basophils (range 4-70) after challenge with anti-IgE Ab. FMLP and C5a led to somewhat lower levels of secretion. A direct comparison of basophils at 88 to 99% purity with basophils from the same preparations, but at lower purities, showed that the amount of IL-4 secretion was proportional to the purity of the basophils. The presence of mRNA for IL-4 (as determined by reverse transcriptase-PCR, competitive reverse transcriptase-PCR, or Northern blots) was also strictly related to the purity of the basophils. IL- 4 mRNA was also found to be constitutively present and was increased after stimulation. The concentration of polyclonal anti-IgE Ab required for optimal IL-4 release was somewhat less than that required for optimal histamine release. IL-4 secretion was slower (t( 1/2 ) of 1.5 hr than histamine release and was inhibited by cycloheximide. In a final series of studies, we found that IL-3 was not required for IL-4 secretion; a short 15-min preincubation with IL-3 resulted in the same or slightly less IL-4 release than no treatment with IL-3. In contrast, an 18-h pretreatment with IL-3 resulted in a nearly tenfold increase in IL-4 secretion. We conclude that human basophils secrete IL-4 in response to several secretagogues and that IL-3 priming is not necessary to observe IL-4 secretion.

Original languageEnglish
Pages (from-to)3006-3016
Number of pages11
JournalJournal of Immunology
Volume152
Issue number6
Publication statusPublished - Mar 15 1994
Externally publishedYes

ASJC Scopus subject areas

  • Immunology

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    MacGlashan, D., White, J. M., Huang, S. K., Ono, S. J., Schroeder, J. T., & Lichtenstein, L. M. (1994). Secretion of IL-4 from human basophils: The relationship between IL-4 mRNA and protein in resting and stimulated basophils. Journal of Immunology, 152(6), 3006-3016.