The relationship between released soluble FceRI-alpha and its cell surface density on human basophils

Abstract

Background The IgE-mediated activation of mast cells and basophils results in the secretion of many substances, including the release of FceRI-alpha subunit. This released alpha subunit can bind IgE and it may act as a down-regulator of subsequent IgE-dependent reactions. However, previous studies do not observe loss of the mass of FceRI-alpha associated with the cells, at least not for human basophils. This study was designed to understand the basis for the discordant observations. Methods Purified human basophils were stimulated with multiple activating secretagogues and supernatants were examined for histamine and released FceRI-alpha. In addition, cell surface IgE densities (occupied and unoccupied) were measured by flow cytometry and total cellular content of mature and immature FceRI-alpha determined with Western blots. Results Released FceRI-alpha, on average, represented 7% of the total surface FceRI before the reaction. The molecular weight of the soluble FceRI-alpha was approximately 54 kD, larger than immature subunit and somewhat smaller than surface subunit. In addition, 1) release ceased long before internalized FceRI-alpha was processed, 2) release was insensitive to Bafilomycin A, 3) release was independent of the starting density of FceRI and 4) release occurred more effectively with non-IgE-dependent stimuli, FMLP or C5a. Conclusions There appears to be relatively constant amount of nearly mature FceRI-alpha that is susceptible to secretion events induced by any form of stimulation. The amount, on average, represents about 7% of the mature form of FceRI-alpha.