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Published online before print July 3, 2007

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(Journal of Leukocyte Biology. 2007;82:1003-1010.)
© 2007 by Society for Leukocyte Biology

Endocytosis, recycling, and degradation of unoccupied FcRI in human basophils

Johns Hopkins Asthma and Allergy Center, Baltimore, Maryland, USA

¹ Correspondence: Johns Hopkins Asthma and Allergy Center, 5501 Hopkins Bayview Circle, Baltimore, MD 21224, USA. E-mail: dmacglas{at}jhmi.edu

Previous studies about basophils and mast cells identified the ability of IgE to up-regulate FcRI expression by a process that depends on stabilization of the surface receptor by IgE. However, the mechanism of loss from the cell surface, when unoccupied, is not known. The current studies have examined whether unoccupied FcRI on basophils is lost by shedding or endocytosis. IgE was dissociated partially from purified human basophils to augment loss of the unoccupied receptor, and comparisons were made between basophils ± IgE resensitization prior to 1-day culture. Incubation did not result in a detectable receptor in culture supernatants. However, in the presence of IL-3, although total cell surface expression decreased by 30% (relative to resensitized cells), FcRI from whole cell lysates was not statistically different between the two conditions. Incubation for 18 h without IL-3 resulted in the same loss from the cell surface but equivalent loss in whole cell lysates. This degradation process was reversible with Bafilomycin A. There was also evidence that the internalized receptor could be recycled. After the initial 18-h down-regulation, the receptor could be found partially restored to the cell surface if IgE were added back to the culture ± cycloheximide. Loss of the unoccupied receptor, as well as accumulation of the receptor under the influence of IgE, was found to be insensitive to the presence of a src-family kinase inhibitor, PP1. These studies establish that the unoccupied receptor is lost by a process of endocytosis, partially recycled to the cell surface, and ultimately degraded by a lysosomal mechanism.

Key Words: mast cells • Fc receptors • allergy

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