Characterization of the D, c, E and G antigens of the Rh blood group system with human monoclonal antibodies.
The human MAbs, anti-D, -c, -E and -G of the Rh blood group system, produced by Epstein-Barr virus transformed B-cell lines, were purified by protein A-Sepharose chromatography and used to characterize the Rh antigens of human red cells. Scatchard plot analyses performed with the radiolabelled MABs indicated that each R2R2 red cell carries 0.43, 0.32 and 0.38 x 10(5) D, c and E binding sites, respectively. About half this number of antigen sites are present on erythrocytes from heterozygote individuals using the appropriate antibody. We found, however, that only 0.18 x 10(5)G antigenic sites were present on each R1R1 red cell. The affinity constants of the anti-D, -E and -G were similar varying from 0.6 to 1.5 x 10(8) M-1 whereas that of the anti-c was much lower (0.035 x 10(8) M-1). The blood group specificity and binding properties indicate that the MAbs behave like the polyclonal anti-Rh reagents. Immunoprecipitation experiments carried out with membranes from R2R2 red cells show that a 30-32 kDa component can be identified whatever the antibody used. The immune complexes involving anti-c, -E or -G antibodies could be formed with the detergent lysates from red cell membranes. In contrast, membrane integrity was a prerequisite for the binding of the anti-D antibodies. Finally, from extraction studies of immunocomplexes with non-ionic detergents it was concluded that all the Rh-active components are bound to the membrane skeleton, suggesting that these molecules may have important function for maintaining red cell shape and viability.
Bloy
C
C
Institut National de Transfusion Sanguine, Paris, France.
Blanchard
D
D
Lambin
P
P
Goossens
D
D
Rouger
P
P
Salmon
C
C
Masouredis
S P
SP
Cartron
J P
JP
D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis*; D004912:Erythrocytes / Q000276:immunology*; D006801:Humans; D007519:Isoantigens / Q000276:immunology; D010455:Peptides / Q000276:immunology; D012204:Rh-Hr Blood-Group System / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007519:Isoantigens; D010455:Peptides; D012204:Rh-Hr Blood-Group System