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International Immunology, Vol. 12, No. 2, 169-176, February 2000
© 2000 Japanese Society for Immunology

Regulation of mast cell signaling through high-affinity IgE receptor by CD45 protein tyrosine phosphatase

Kiichi Murakami, Shintaro Sato, Shigeharu Nagasawa and Toshiyuki Yamashita

Department of Hygienic Chemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita-ku, Sapporo 060-0812, Japan

Correspondence to: T. Yamashita

The transmembrane tyrosine phosphatase CD45 regulates the activity of src family protein tyrosine kinases (PTK) and thereby influences the signaling via such receptors as T and B cell antigen receptors associated with these PTK. However, its implication in signaling through the mast cell receptor with high affinity for IgE (Fc{epsilon}RI) is less clear, although Lyn, a member of the src family, plays an important role in Fc{epsilon}RI-mediated signaling. To define a role for CD45 in Fc{epsilon}RI signal transduction, we established CD45 high expressing rat basophilic leukemia cell lines (RBL-CD45H) and cell lines expressing trace amounts of CD45 (RBL-CD45L). We demonstrate that although all RBL-CD45L cell lines degranulate following IgE- and antigen-induced Fc{epsilon}RI aggregation, the response is significantly reduced at a low dose of antigen. The cells show a delayed and slowed Ca2+ mobilization even though at a higher dose where the cells degranulate to a similar extent as RBL-CD45H. This diminished Ca2+ response is restored by reconstitution of RBL-CD45L with a chimeric molecule containing the cytoplasmic phosphatase domains of rat CD45. Furthermore, tyrosine phosphorylation of Fc{epsilon}RI, association of Fc{epsilon}RI with Lyn and PTK activity associated with Fc{epsilon}RI, all of which are enhanced upon Fc{epsilon}RI aggregation in RBL-CD45H, are impaired in RBL-CD45L. Finally, we show that Fc{epsilon}RI is physically associated with CD45 in RBL-CD45H prior to receptor aggregation. Thus, we propose that, although not indispensable in mast cell degranulation, CD45 positively regulates the signaling through Fc{epsilon}RI by promoting the activation of Fc{epsilon}RI-associated Lyn.

Keywords: Fc{varepsilon}RI, Lyn, tyrosine phosphorylation, rat basophilic leukemia

Transmitting editor: D. Kitamura


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