Neutrophil activation through high-affinity Fc gamma receptor using a monomeric antibody with unique properties.
The high-affinity, type I Ig Fc receptor (Fc gamma RI) for human IgG1, human IgG3, murine IgG2a, and murine IgG3 is highly expressed on monocytes, neutrophils (PMN) in certain disease states, and phagocytes treated with interferon-gamma (IFN-gamma). We studied the activation of the human PMN oxidative burst and stimulated fluid pinocytosis induced by three monoclonal antibodies (MoAbs) directed against Fc gamma RI (CD64) to study the role of this receptor in Fc-mediated cellular activation. All three MoAbs were capable of triggering PMN activation from IFN-gamma-treated PMN when cross-linked with goat antimurine Ig reagents. However, MoAb 197 alone demonstrated a concentration-dependent activation of the oxidative burst without the use of a second cross-linking antibody. The oxidative burst and stimulated fluid pinocytosis responses induced by monomeric MoAb 197 could be elicited only after the IFN-gamma induction of approximately 8,000 Fc gamma RI receptor equivalents and did not occur in freshly isolated or control-cultured PMN. Competitive blocking of Fc binding of MoAb 197 by human IgG or purified Fc fragments inhibited cellular activation. We believe the ability of MoAb 197 to activate these oxidative burst and fluid pinocytic responses was because of its murine IgG2a subclass, which allowed it to function as a trivalent anti-Fc gamma RI antibody binding through the combination of its two FAB regions and the Fc domain. This study demonstrates that the cross-linking of CD64 can activate PMN oxidative and endocytic responses and supports a role for Fc gamma RI in the human neutrophil inflammatory response.
Akerley
W L
WL
Department of Pathology, Dartmouth-Hitchcock Medical Center, Hanover, NH 03756.
Guyre
P M
PM
Davis
B H
BH
D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D000939:Epitopes / Q000276:immunology; D005455:Fluorescent Antibody Technique; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000378:metabolism; D007700:Kinetics; D009504:Neutrophils / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D010084:Oxidation-Reduction; D010873:Pinocytosis / Q000187:drug effects / Q000502:physiology; D011961:Receptors, Fc / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007141:Immunoglobulin Fc Fragments; D011961:Receptors, Fc