低親和性IgGレセプター(FcγRII)を介するヒト好塩基球の活性化機序に関する研究
フローサイトメーターによるカルシウム動態の解析
1994 年 106 巻 7-8 号 p. 847-860
詳細
1994 年 106 巻 7-8 号 p. 847-860
A new method for flow cytometric analysis of calcium mobilization in human peripheral blood basophils without prior purification was developed. The method is based on dual color analysis of centrifugation-enriched mononuclear cell populations using fluo-3 and phycoerythrin (PE)-conjugated CD2, CD14, CD16, CD19 monoclonal antibodies (mAb) to stain contaminated cells. This technique allows the detection of fluo-3 fluorescence as a measure of an increase in the cytoplasmic free calcium concentration ( [Ca2+] 1) while simultaneously discriminating PE-mAb-unlabelled basophils.
To clarify whether the human peripheral blood basophil is activated through the low affinity IgG receptor, Fc γ RII, as well as the high affinity IgE receptor, Fc ε R I, calcium mobilization after Fc γ RII stimulation was analyzed by this method. After cross-linking of Fc γ RII, transient [Ca2+] 1 elevation was observed but there was no apparent difference with interleukin-3 (IL-3)-treated cells, and no significant histamine release was observed with or without short pre-incubation of IL-3. These findings suggest that the cross-linking of Fc γ RII, not only Fc ε R I, can activate human basophils which may result in mediator release other than histamine.
