LIPOSOME-MEDIATED DELIVERY OF DEOXYRIBONUCLEIC-ACID TO CELLS - ENHANCED EFFICIENCY OF DELIVERY RELATED TO LIPID-COMPOSITION AND INCUBATION CONDITIONS
LIPOSOME-MEDIATED DELIVERY OF DEOXYRIBONUCLEIC-ACID TO CELLS - ENHANCED EFFICIENCY OF DELIVERY RELATED TO LIPID-COMPOSITION AND INCUBATION CONDITIONS
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DOI：
10.1021/bi00527a031
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发表时间：
1981-01-01
期刊：
影响因子：
2.9
通讯作者：
PAPAHADJOPOULOS, D
中科院分区：
文献类型：
--
作者：
FRALEY, R;STRAUBINGER, RM;PAPAHADJOPOULOS, D
Delivery of liposome-encapsulated SV40 DNA to African green monkey cells was used as a probe to study liposome-cell interactions and to determine conditions which favor the intracellular delivery of liposome contents to cells. The efficiency of DNA delivery by various liposome preparations (monitored by infectivity assays) was dependent on the magnitude of vesicle binding to cells and on the resistance of liposomes to cell-induced leakage of contents. Acidic phospholipids were more effective in binding and delivery and phosphatidylserine (PS) was best in both aspects. The inclusion of 50 mol % cholesterol in liposomes reduced the cell-induced leakage of vesicle contents (2- to 5-fold) and substantially enhanced the delivery of DNA to cells (2- to 10-fold). Following incubation of cells with negatively charged liposomes containing SV40 DNA, infectivity was enhanced greatly by brief exposure of the cells to glycerol solutions. Only slight enhancement by glycerol was observed for SV40 DNA encapsulated in neutral or positively charged liposomes. The results of competition experiments between empty phosphatidylcholine liposomes and DNA-containing PS liposomes suggested possible differences in the interaction of neutral and negatively charged liposome preparations with cells. Morphological studies indicated that glycerol treatment stimulates membrane ruffling and vacuolization and that the enhanced uptake of liposomes occurs by an endocytosis-like process. Results obtained with metabolic inhibitors were also consistent with the interpretation that the enhancement of liposome delivery in glycerol-treated cells occurs via an energy-dependent endocytotic pathway. Pretreatment of cells with chloroquine, a drug which alters lysosomal activity, further enhanced infectivity in glycerol-treated cells (4-fold). The involvement of a lysosomal processing step at some point in the expression of liposome-encapsulated DNA is suggested and illustrates the possibility of altering cellular metabolism to engineer more efficient delivery by liposomes. Under optimal conditions, the efficiency of liposome-mediated SV40 DNA delivery was increased > 1000-fold over that obtained by simply incubating cells with liposomes. These conditions may enhance delivery of other molecules, besides DNA, which are encapsulated in liposomes.
