Complement-opsonized IgG antibody/dsDNA immune complexes bind to CR1 clusters on isolated human erythrocytes.
We used fluorescence microscopy, quantitative FACS analyses, and radioimmunoassays to examine the distribution of complement (C3b)-opsonized antibody/dsDNA immune complexes (IC) bound to normal human erythrocytes (RBCs) via immune adherence (IA). IC were detected with fluorescent anti-human IgG, and the RBC IA-receptor (CR1) was revealed with monoclonal antibodies (Mabs) to CR1 and fluorescent anti-mouse IgG. Under saturating conditions RBCs exhibit a large heterogeneity in binding; a significant fraction binds no IC. The positions of bound IC coincide with CR1 clusters on RBCs, as predicted by several investigators. FACS experiments indicate an excellent correlation between CR1 number and IC binding within an RBC population. The number of CR1 clusters able to bind IC is proportional to the number of CR1 per RBC. However, IC (fluorescent spots) detected per RBC (on average less than 10) are less than the average number of IC bound (20-30). This suggests that each fluorescent spot represents a small number of aggregated IC bound to a CR1 cluster. These "patches" of aggregated complexes may facilitate transfer of RBC-bound IC to cells of the mononuclear phagocytic system. Finally, not all the CR1 clusters bind IC, suggesting that proper geometric alignment of multiple C3b per IC with several CR1 in the cluster is required for IA.
Taylor
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RP
Department of Biochemistry, University of Virginia School of Medicine, Charlottesville 22908.
Pocanic
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F
Reist
C
C
Wright
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EL
D000936:Antigen-Antibody Complex / Q000032:analysis* / Q000378:metabolism; D003179:Complement C3b / Q000276:immunology*; D003165:Complement System Proteins / Q000276:immunology*; D004247:DNA / Q000276:immunology*; D004912:Erythrocytes / Q000737:chemistry*; D005434:Flow Cytometry; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis*; D008856:Microscopy, Fluorescence; D010587:Phagocytosis; D011951:Receptors, Complement / Q000032:analysis*; D017463:Receptors, Complement 3b
D000936:Antigen-Antibody Complex; D007074:Immunoglobulin G; D011951:Receptors, Complement; D017463:Receptors, Complement 3b; D003179:Complement C3b; D003165:Complement System Proteins; D004247:DNA