A critical role for PI 3-kinase in cytokine-induced Fcα-receptor activation

Abstract

AbstractFc-receptors, such as FcαR and FcγRII, play an important role in leukocyte activation, and rapid modulation of ligand binding (“activation”) is critical for receptor regulation. We have previously demonstrated that ligand binding to Fc-receptors on human eosinophils is dependent on cytokine stimulation. Utilization of pharmacological inhibitors provided evidence that the phenomenon of interleukin (IL)-5 induced immunoglobulin A (IgA) binding to human eosinophils requires activation of phosphatidylinositol 3-kinase (PI3K). However, eosinophils are refractory to manipulation by molecular techniques such as DNA transfection or viral infection. Here we utilize an IL-3 dependent pre-B cell line to investigate the molecular mechanism of cytokine-mediated ligand binding to FcαR. In this system, IgA binding is dependent on IL-3, similarly to the requirement for IL-5 of eosinophils. We show that IL-3-mediated activation of FcαR (CD89) requires the activation of PI3K, independent of p21ras activation. Co-expression of dominant negative (▵p85) and active (p110_K227E) forms of PI3K demonstrate that the affinity switch regulating FcαR activation requires PI3K. Moreover, overexpression of PI3K is both necessary and sufficient for activation of FcαR. Furthermore, we show that IL-3/IL-5/GM-CSF induced inside-out signaling pathways activating FcαR require the involvement of protein kinase C downstream of PI3K. Finally, we show that these inside-out signaling pathways responsible for Fcα-receptor modulation require CD89, independent of its association with the FcRγ chain.