IgE-Dependent expression of mRNA for IL-4 and IL-5 in human lung mast cells

Yoshimichi Okayama, Corinne Petit-Frére, Olivier Kassel, Amanda Semper, Diana Quint, Manuel J. Tunon-de-Lara, Peter Bradding, Stephen T. Holgate, Martin K. Church*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

143 Citations (Scopus)

Abstract

By using the reverse transcriptase (RT)-PCR and in situ hybridization we have studied the expression of mRNA for IL-5 and IL-4 in human lung mast cells induced by cross-linkage of high affinity FcεRs. Lung mast cells were purified using affinity magnetic selection with mAb YB5.B8 against c-kit to achieve a final mast cell purity >93%. Purified mast cells were precultured with stem cell factor (SCF) (10 ng/ml) and myeloma IgE (3 μg/ml) for 16 h before challenge with anti-IgE (1 or 10 μg/ml). IgE-dependent activation of lung mast cells caused expression of IL-5 mRNA, which was evident by 2 h and persisted for up to 48~72 h in all of 12 experiments, whereas IL-4 mRNA expression was of a shorter duration and was demonstrable in 6 of 13 experiments. We confirmed that mast cells, and not T cells, were the source of these cytokine messages by using reverse transcriptase-PCR in cell preparations containing known numbers of mast cells and T cells, in situ hybridization in enriched mast cell preparations, and double in situ hybridization-immunocytochemical staining. IL-5 mRNA expression did not require the pretreatment of cells with SCF, whereas expression of IL-4 mRNA seemed to require both anti-IgE and SCF. The strength of IL-5 mRNA signal was related to anti-IgE concentration. Immunoreactive IL-5 was detectable 8 h after anti-IgE challenge, and 106 mast cells generated a mean of 731 ± 400 pg of IL-5 into the supernatant during 48-h culture, but no IL-4 product was detectable. These findings demonstrate the capacity of human lung mast cells to transcribe IL-4 and IL-5 after IgE-dependent activation and to synthesize and release immunoreactive IL-5.

Original languageEnglish
Pages (from-to)1796-1808
Number of pages13
JournalJournal of Immunology
Volume155
Issue number4
Publication statusPublished - 1995
Externally publishedYes

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