Reciprocal regulation of cultured human mast cell cytokine production by IL-4 and IFN-gamma.

Abstract

T(H)1 and T(H)2 cytokines are thought to regulate allergic inflammation. Two key regulatory cytokines, IL-4 and IFN-gamma, were examined for their effects on cytokine production by cultured human mast cells (CHMCs). CHMCs were obtained by culturing cord blood-derived CD34(+) cells in the presence of stem cell factor and IL-6 for 14 to 16 weeks. CHMCs were passively sensitized with human myeloma IgE and supplemented with or without IL-4 or IFN-gamma. After the sensitization, CHMCs were stimulated with anti-FcepsilonRIalpha mAb. Concentrations of secreted cytokines were measured by using ELISA, and cytokine messenger RNA was analyzed by using quantitative competitive RT-PCR. IL-4 profoundly enhanced FcepsilonRI-mediated production of macrophage inflammatory protein (MIP) 1alpha, IL-8, and GM-CSF. For example, the enhancement by IL-4 (10 ng/mL) of the production of MIP-1alpha, IL-8, and GM-CSF was 25-, 7-, and 90-fold, respectively, after 6 hours. IL-4 also enhanced levels of FcepsilonRI-induced cytokine messenger RNA but to a lesser degree. In contrast, IFN-gamma inhibited FcepsilonRI-induced production of MIP-1alpha, IL-8, and GM-CSF. For example, the inhibition by IFN-gamma (10 ng/mL) of FcepsilonRI-mediated production of MIP-1alpha, IL-8, and GM-CSF was 80%, 75%, and 95%, respectively. IFN-gamma also suppressed FcepsilonRI-induced messenger RNA expression of these cytokines. Neither IL-4 nor IFN-gamma affected the kinetics of cytokine production by CHMCs. These data suggest that IL-4 and IFN-gamma may influence allergic reactions by modulating human mast cell cytokine production.