A critical role for PI 3-kinase in cytokine-induced Fcalpha-receptor activation.
Fc-receptors, such as FcalphaR and FcgammaRII, 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 FcalphaR. 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 FcalphaR (CD89) requires the activation of PI3K, independent of p21ras activation. Co-expression of dominant negative (triangle upp85) and active (p110_K227E) forms of PI3K demonstrate that the affinity switch regulating FcalphaR activation requires PI3K. Moreover, overexpression of PI3K is both necessary and sufficient for activation of FcalphaR. Furthermore, we show that IL-3/IL-5/GM-CSF induced inside-out signaling pathways activating FcalphaR require the involvement of protein kinase C downstream of PI3K. Finally, we show that these inside-out signaling pathways responsible for Fcalpha-receptor modulation require CD89, independent of its association with the FcRgamma chain. (Blood. 2000;95:2037-2043)
Bracke
M
M
Department of Pulmonary Diseases, University Medical Center, Utrecht, The Netherlands.
Nijhuis
E
E
Lammers
J W
JW
Coffer
P J
PJ
Koenderman
L
L
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism*; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000378:metabolism; D006801:Humans; D007070:Immunoglobulin A / Q000378:metabolism; D007377:Interleukin-3 / Q000378:metabolism; D015848:Interleukin-5 / Q000378:metabolism; D008024:Ligands; D051379:Mice; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases; D019869:Phosphatidylinositol 3-Kinases / Q000502:physiology*; D011485:Protein Binding; D011493:Protein Kinase C / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt; D011961:Receptors, Fc / Q000378:metabolism*; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection; D048051:p38 Mitogen-Activated Protein Kinases
D015703:Antigens, CD; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D016207:Cytokines; D004791:Enzyme Inhibitors; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D007070:Immunoglobulin A; D007377:Interleukin-3; D015848:Interleukin-5; D008024:Ligands; D019950:Mitogen-Activated Protein Kinase 1; D020928:Mitogen-Activated Protein Kinases; D019869:Phosphatidylinositol 3-Kinases; D011493:Protein Kinase C; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D051057:Proto-Oncogene Proteins c-akt; D011961:Receptors, Fc; D048051:p38 Mitogen-Activated Protein Kinases; C036832:Fc(alpha) receptor