Immune complexes formed following the binding of anti-platelet factor 4 (CXCL4) antibodies to CXCL4 stimulate human neutrophil activation and cell adhesion.
We tested the possibility that immune complexes formed following platelet factor 4 (PF4/CXCL4) binding to anti-PF4 antibody can stimulate neutrophil activation, similar to previous reports with platelets. Monoclonal Abs against PF4 and IgG from a heparin-induced thrombocytopenia (HIT) patient were applied. We observed that although PF4 or anti-PF4 antibody alone did not alter neutrophil function, costimulation with both reagents resulted in approximately 3-fold increase in cell surface Mac-1 expression, enhanced cell adhesion via L-selectin and CD18 integrins, and degranulation of secondary and tertiary granules. The level of Mac-1 up-regulation peaked at an intermediate PF4 dose, suggesting that functional response varies with antigen-antibody stoichiometry. PF4 binding to neutrophils was blocked by chondroitinase ABC. Cell activation was inhibited by both chondroitinase ABC and anti-CD32/FcgammaRII blocking mAb, IV.3. Confocal microscopy demonstrated that immune complexes colocalize with CD32a. Studies with HIT IgG demonstrated that neutrophils could be activated in the absence of exogenous heparin. These data, together, show that leukocyte surface chondroitin sulfates promote neutrophil activation by enhancing immune-complex binding to CD32a. Studies with recombinant PF4 suggest a role for arginine 49 in stabilizing PF4-chondroitin binding. Neutrophils activated via this mechanism may contribute to thrombosis and inflammation in patients mounting an immune response to PF4-heparin.
Xiao
Zhihua
Z
Chemical and Biological Engineering, State University of New York,Buffalo, NY 14260, USA.
Visentin
Gian P
GP
Dayananda
Kannayakanahalli M
KM
Neelamegham
Sriram
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000936:Antigen-Antibody Complex / Q000096:biosynthesis / Q000502:physiology*; D000937:Antigen-Antibody Reactions; D002448:Cell Adhesion / Q000276:immunology*; D002809:Chondroitin Sulfates / Q000378:metabolism; D006493:Heparin / Q000009:adverse effects; D006801:Humans; D007962:Leukocytes / Q000737:chemistry; D018375:Neutrophil Activation / Q000276:immunology*; D010978:Platelet Factor 4 / Q000276:immunology*; D051381:Rats; D017452:Receptors, IgG / Q000378:metabolism*; D013921:Thrombocytopenia / Q000139:chemically induced / Q000276:immunology
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; C080568:Fc gamma receptor IIA; D017452:Receptors, IgG; D010978:Platelet Factor 4; D006493:Heparin; D002809:Chondroitin Sulfates