Haptenic activity of galactosyl ceramide and its topographical distribution on liposomal membranes. I. Effect of cholesterol incorporation.
The relation between the immune-reaction of phosphatidylcholine liposomes containing spin-labeled galactosyl ceramide with or without cholesterol and the topographical distribution of the glycolipid in membranes was studied. In egg yolk phosphatidylcholine liposomes, both immune agglutination and antibody binding occurred, irrespectively of the presence of cholesterol, though the motion of the fatty acyl chain of spin-labeled galactosyl ceramide was restricted by cholesterol. In dipalmitoyl phosphatidylcholine liposomes, unlike in egg yolk phosphatidylcholine liposomes, the immune-reaction depended on the cholesterol content. The electron spin resonance (ESR) spectra of spin-labeled galactosyl ceramide in dipalmitoyl phosphatidylcholine liposomes indicated that cholesterol affected the topographical distribution of spin-labeled galactosyl ceramide in the liposomes. Without cholesterol, most of the spin-labeled galactosyl ceramide was clustered on the dipalmitoyl phosphatidylcholine membrane, but with increase of cholesterol, random distribution of hapten on the membrane increased. The cholesterol-dependent change in the topographical distribution of hapten on the membranes was parallel with that of immune reactivity. 'Aggregates' composed solely of galactosyl ceramide did not show any binding activity with antibody. The findings suggest that the recognition of galactosyl ceramide by antibody depended on the topographical distribution of hapten molecules. Phosphatidylcholine and/or cholesterol may play roles as 'spacers' for the proper distribution of 'active' haptens on the membranes. The optimum density of haptens properly distributed on liposomal membranes is discussed.
D000371:Agglutination; D000818:Animals; D000936:Antigen-Antibody Complex; D002554:Cerebrosides; D002784:Cholesterol; D004578:Electron Spin Resonance Spectroscopy; D005699:Galactosylceramides; D006241:Haptens; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D008081:Liposomes; D010713:Phosphatidylcholines / Q000276:immunology*; D013113:Spin Labels
D000936:Antigen-Antibody Complex; D002554:Cerebrosides; D005699:Galactosylceramides; D006241:Haptens; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D008081:Liposomes; D010713:Phosphatidylcholines; D013113:Spin Labels; D002784:Cholesterol