Interactions of PC/Chol and PS/Chol liposomes with human cells in vitro

Interactions between phosphatidylcholine (PC) or phosphatidylserine (PS) liposomes and human umbilical vein endothelial cells (HUVEC) or human promyelocytic leukemia cells (HL60) were investigated. Pyramine encapsulating or rhodamine incorporating small unilamellar liposomes with mean diameters around 80 nm (demonstrated to retain encapsulated material and to be nontoxic under experimental conditions) were used. Liposome uptake by both types of cells increased when increasing amounts of vesicles were co-incubated. For both lipid compositions, the interaction with HUVEC was very fast (association reached a plateau within 5 min) and so was the release of internalized vesicles (90% within 10 min at 37 degrees C). The reduced association values at 4 degrees C and the punctuate fluorescence observed in the cell cytoplasm after interaction, were indicative of whole liposome internalization. This internalization was clathrin-independent, since it was not inhibited by sodium azide and deoxyglucose. Pre-treatment of HUVEC with filipin or NEM resulted in modification of the interaction, something that could be due to alterations in the biochemical characteristics of HUVEC membranes that inhibit vesicular processes. In HL-60 cells, a slower association and faster release of PC/Chol liposomes was demonstrated, while association of both liposomes with these cells was energy-and temperature-independent. Nevertheless, morphological studies revealed differences in the interactions: A bright fluorescent rim observed after interaction with PC/Chol liposomes, suggests that these liposomes were adsorbed on the surface of HL60 cells, while the uniform cytoplasmic fluorescence observed after incubation with PS/Chol liposomes was indicative of fusion as the interaction mechanism.