lysophosphatidylethanolamine and 1-2-dioleoyloxy-3-(trimethylammonium)propane

The effect of curvature stress on the efficiency of cationic liposome-induced fusion between rabbit erythrocytes was studied. Multilamellar cationic liposomes containing 1,2-dioleoyl-3-trimethylammoniumpropane (DOTAP) and different PEs (1,2-dilnoleoyl-sn-glycero-3-phosphoethanolamine (dilin-PE), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), and lysophosphatidylethanolamine, egg (lyso-PE)) were used to induce cell-cell fusion. It was found that high cell-cell fusion yield (FY) of about 50% could be achieved in sucrose solution by using cationic liposomes containing 50% DOTAP. Cell-cell fusion was assayed by shape criterion and was verified by fluorescence microscopy, using a membrane dye mixing method. The curvature stress, as a result of mixing unsaturated PEs in cationic liposomes, had a significant effect on cell-cell FY which increased with the degree of acyl chain unsaturation, in the order dilin-PE > DOPE > POPE > lyso-PE. Replacement of dilin-PE, DOPE, or POPE by lyso-PE gradually in cationic liposomes lowered the cell-cell FY from 50% to 1%. Furthermore, cationic liposome induced cell lysis, and fusion between cationic liposomes and cells, as assayed by the N-(lissamine rhodamine B sulfonyl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt and N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-1,2- dihexadecanoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt (Rh-PE/NBD-PE) energy transfer method, followed the same order as that for cell-cell fusion. Fusion between the negatively charged PS liposomes and cationic liposomes also followed the same order. The electric double layer screening by electrolytes in NaCl-containing solution and phosphate buffered saline (PBS) was found to reduce the cell-cell FY and cell lysis. These findings suggest that liposome-induced cell-cell fusion was achieved by cationic liposomes serving as fusion-bridges among cells.

Topics: Animals; Cations; Cell Fusion; Erythrocytes; Fatty Acids, Monounsaturated; In Vitro Techniques; Lipids; Liposomes; Lysophospholipids; Membrane Fusion; Phosphatidylethanolamines; Quaternary Ammonium Compounds; Rabbits; Static Electricity