Effect of membrane cholesterol depletion on the immunoreactivity of the D antigen and IgG anti-D.

The immunoreactivity of the main Rh antigen (D) and its corresponding antibody, as determined by a ti-IgG to IgG combining ratio, antibody dissociation and antigen accessibility to antibody, was examined in cholesterol depleted human red cells and ghost membranes. The anti-IgG reactivity of IgG anti-D bound to cholesterol depleted red cells and ghosts was demonstrably enhanced in vitro and in electron microscopy studies, particularly in ghosts. Dissociation of cell bound anti-D during buffer incubation was greater after cholesterol depletion, especially in ghosts. There was also reduced binding of anti-D to cholesterol-depleted cells as previously reported. All these effects appeared to be independent of endogenous or exogenous proteolysis in either cholesterol-depleted membranes or controls as judged from membrane electrophoretic analyses. A2C, an agent which increases membrane fluidity, had no effect on anti-D binding or the antiglobulin reactivity of cell bound IgG. A reduction in anti-D binding also was observed in red cells depleted of cholesterol following immobilization of membrane proteins by glutaraldehyde crosslinking. The findings show that cholesterol depletion not only affects the antigen but also Rh antibody reactivity. They also suggest that factors other than vertical antigen movement in a fluid bilayer may influence the behavior of the D antigen in cholesterol-modified erythrocytes.