CR3 (alphaM beta2; CD11b/CD18) restores IgG-dependent phagocytosis in transfectants expressing a phagocytosis-defective Fc gammaRIIA (CD32) tail-minus mutant.
Previous studies have suggested that complement receptors cooperate with Fc receptors to mediate Ab-dependent effector functions. In the present study, we tested the capacity of complement receptor type 3 (CR3; alphaM beta2; CD11b/CD18) to participate in Fc gammaRIIA (CD32)-mediated phagocytosis. To test this hypothesis, we transfected a phagocytosis-defective tail-minus mutant of Fc gammaRIIA into fibroblasts that did or did not express CR3. As controls, cells exposed to the transfection protocol but not expressing either CR3 or Fc gammaRIIA constructs were also selected. Expression of cell surface receptors was confirmed by both flow cytometry and fluorescence microscopy. Cells were tested for their ability to bind and internalize IgG-opsonized erythrocytes and for surface proximity of CR3 and Fc gammaRIIA using resonance energy transfer techniques. Resonance energy transfer studies showed a physical proximity between CR3 and Fc gammaRIIA on cell surfaces. Cells expressing only the tail-minus mutant of Fc gammaRIIA were unable to internalize erythrocytes but showed significant binding ability. In contrast, cells expressing both mutant Fc gammaRIIA and CR3 internalized opsonized erythrocytes. These results show that CR3 can complement the phagocytic function of defective Fc gammaRIIA. These data suggest that CR3 is capable of transducing a phagocytic signal generated by Fc gammaRIIA recognition events that mediates internalization of IgG-opsonized targets.
Worth
R G
RG
Department of Biological Sciences, Wayne State University, Detroit, MI 48202, USA.
Mayo-Bond
L
L
van de Winkel
J G
JG
Todd
R F
RF
Petty
H R
HR
D016475:3T3 Cells; D000818:Animals; D000936:Antigen-Antibody Complex / Q000378:metabolism; D005816:Genetic Complementation Test; D007074:Immunoglobulin G / Q000502:physiology; D016177:Macrophage-1 Antigen / Q000502:physiology*; D051379:Mice; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence / Q000379:methods; D010587:Phagocytosis; D011940:Receptor Aggregation; D017452:Receptors, IgG / Q000502:physiology*; D013329:Structure-Activity Relationship; D014162:Transfection
D000936:Antigen-Antibody Complex; D007074:Immunoglobulin G; D016177:Macrophage-1 Antigen; D017452:Receptors, IgG