Analysis of C3-receptor activity on human B-lymphocytes and isolation of the complement receptor type 2 (CR2).
The rosetting of defined C3-fragment-coated sheep erythrocytes to B-cell-enriched tonsil lymphocytes was measured. The rosetting lymphocytes were homogeneous with respect to expression of C3b, iC3b and C3d receptors. Isolation of receptors for C3 fragments from surface-radioiodinated lymphocytes by affinity chromatography on immobilized C3u, iC3b and C3d,g produced two proteins with partially overlapping specificities. A protein of 240 000 Mr, recognized by the monoclonal antibody To5 and identified as CR1 (complement receptor type 1), had affinity for C3u and iC3b. A protein of 145 000 Mr, recognized by the monoclonal antibody B2, had affinity for all three C3 fragments. Inhibition of rosetting by antibodies to these proteins indicates that CR1 is responsible for C3b-mediated rosetting and that the 145000-Mr receptor (CR2) is responsible for C3d-mediated rosetting. Partial inhibition by both anti-CR1 and anti-CR2 antibodies of iC3b-mediated rosetting indicates that both receptors are involved in iC3b-mediated rosetting. No other protein appears to be involved in tonsil B-cell rosetting to C3-fragment-coated cells. A method for preparing CR2 from tonsil lymphocytes based on affinity chromatography on C3d,g-Sepharose has been developed. Forty tonsil pairs (2 X 10(10) B-cells) yield about 40 micrograms of pure protein equivalent to a purification of 6500-fold from a detergent extract.
D000906:Antibodies / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology*; D002352:Carrier Proteins / Q000276:immunology; D011232:Chemical Precipitation; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D006801:Humans; D016177:Macrophage-1 Antigen; D008565:Membrane Proteins / Q000276:immunology; D014066:Palatine Tonsil / Q000166:cytology; D011951:Receptors, Complement / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification*; D017464:Receptors, Complement 3d; D012397:Rosette Formation
D000906:Antibodies; D002352:Carrier Proteins; D016177:Macrophage-1 Antigen; D008565:Membrane Proteins; D011951:Receptors, Complement; D017464:Receptors, Complement 3d