Neutrophil activation by anti-beta 2 glycoprotein I monoclonal antibodies via Fc gamma receptor II.
Murine monoclonal antibodies (mAbs) to human beta 2-glycoprotein I (beta 2GPI), a plasma protein required for the binding of some antiphospholipid antibodies, have been shown to possess lupus anticoagulant properties and to activate platelets via Fc gamma receptor (Fc gamma R) crosslinking. Here we investigated their ability to induce polymorphonuclear leukocyte (PMN) functional responses. The six mAbs (IgG1 isotype) tested in combination with beta 2GPI led to a concentration-dependent activation of human PMNs as appreciated by granule release, H2O2 production, and cytosolic Ca2+ increase. This activation process was accompanied by the enhancement of PMN-mediated heparan sulfate loss from the endothelial cell line EA.hy 926 without evidence for cell lysis or detachment. F(ab')2 fragments of one of the mAbs bound to PMNs in a beta 2GPI-dependent manner but were devoid of activating effects. Carbamylated beta 2GPI was unable to mediate PMN-antibody binding and subsequent activation. In addition, cationization of beta 2GPI or removal of its sialic acid groups led to higher efficiency in binding to the PMN surface and triggering activation in comparison with the untreated protein. Thus, the process of PMN activation depends on mAb binding to these cells through both Fab (via beta 2GPI) and Fc domains, as confirmed by the suppression of all responses upon treatment with an anti-Fc gamma RII, but not anti-Fc gamma RIII, antibody. Our data suggest a model of cellular activation by beta 2GPI-dependent antiphospholipid antibodies.
Arvieux
J
J
Laboratoire d'Immunologie, INSERM U238, Grenoble, France.
Jacob
M C
MC
Roussel
B
B
Bensa
J C
JC
Colomb
M G
MG
D000911:Antibodies, Monoclonal / Q000276:immunology*; D002118:Calcium / Q000378:metabolism; D015550:Cell Degranulation; D003571:Cytochalasin B / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000378:metabolism; D006023:Glycoproteins / Q000276:immunology*; D006497:Heparan Sulfate / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D066298:In Vitro Techniques; D018375:Neutrophil Activation; D009504:Neutrophils / Q000502:physiology*; D010196:Pancreatic Elastase / Q000378:metabolism; D017452:Receptors, IgG / Q000276:immunology; D016897:Respiratory Burst; D053482:beta 2-Glycoprotein I
D000911:Antibodies, Monoclonal; D002118:Calcium; D003571:Cytochalasin B; D006023:Glycoproteins; D006497:Heparan Sulfate; D006861:Hydrogen Peroxide; D010196:Pancreatic Elastase; D017452:Receptors, IgG; D053482:beta 2-Glycoprotein I