Influence of vesicle size on complement-dependent immune damage to liposomes.
Complement-dependent antibody-mediated damage to multilamellar lipid vesicles (MLVs) normally results in a maximum release of 50-60% of trapped aqueous marker. The most widely accepted explanation for this is that only the outermost lamellae of MLVs are attacked by complement. To test this hypothesis, complement damage to two different types of large unilamellar vesicles (LUVs), large unilamellar vesicles prepared by the reverse-phase evaporation procedure (REVs) and large unilamellar vesicles prepared by extrusion techniques (LUVETs), were determined. In the presence of excess antibody and complement the LUVs released a maximum of only approx. 25 to 40% of trapped aqueous marker, instead of close to 100% that would be expected. Since small unilamellar vesicles apparently differ from LUVs in that they can release 100% of trapped aqueous marker it appeared that the size of the vesicles was an important factor. Because of these observations the influence of MLV size on marker release was examined. Three populations of MLVs of different sizes were separated by a fluorescence activated cell sorter. Assays of the separated MLV populations showed that the degree of complement-dependent marker release was inversely related to MLV size. No detectable glucose was taken up by MLVs when glucose was present only outside the liposomes during complement lysis. Our results can all be explained by the closing, or loss, of complement channels. We conclude that complement channels are only transiently open in liposomes, and that loss of channel patency may be due to either channel closing or to loss of channels.
D000818:Animals; D000920:Antibody-Dependent Cell Cytotoxicity; D000936:Antigen-Antibody Complex; D002417:Cattle; D002554:Cerebrosides; D002784:Cholesterol; D003165:Complement System Proteins / Q000276:immunology*; D004134:Dimyristoylphosphatidylcholine; D007106:Immune Sera; D008081:Liposomes; D011663:Pulmonary Surfactants; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship
D000936:Antigen-Antibody Complex; D002554:Cerebrosides; D007106:Immune Sera; D008081:Liposomes; D011663:Pulmonary Surfactants; D003165:Complement System Proteins; D002784:Cholesterol; D004134:Dimyristoylphosphatidylcholine