Cultured Human Mast Cells Are Heterogeneous for Expression of the High-Affinity IgE Receptor FcεRI

Objective: We determined the density of Fc epsilon RI on mast cells cultured from cord (CBMC) and peripheral blood (PBMC) and studied the kinetics of the response through Fc epsilon RI. Methods: Mast cells were cultured from CD133+ progenitors from peripheral or cord blood. Fc epsilon RI was stabilized by culture with 2 mu g/ml IgE. Cells were activated by addition of anti-Fc epsilon RI antibody (1 ng/ml-10 mu g/ml). Maximal activation, sensitivity, and cooperativity were determined. Results: All cultures were homogeneous for tryptase and metachromasy. All cells expressing Fc epsilon RI could be activated by cross-linking Fc epsilon RI to upregulate CD63. PBMC bind 203,000 molecules of IgE/cell. Stabilization of Fc epsilon RI with IgE doubled the number of CD63+ cells (p = 0.0001) and increased the sensitivity (from 0.083 to 0.013 mu g/ml anti-Fc epsilon RI) and the slope factor (from 10.8 to 68) of PBMC but not of CBMC. Anti-IgE reversed these effects (p = 0.0002) but did not reduce activation levels below that of cell lines not stabilized with IgE. Conclusion: Baseline expression of Fc epsilon RI is independent of anti-IgE. The fraction of PBMC that binds high levels of IgE can be activated through Fc epsilon RI. Copyright (C) 2011 S. Karger AG, Basel