Hodgson C P, Solaiman F
Creighton Cancer Center, Dept. of Biomedical Sciences, Creighton School of Medicine, Omaha, NE 68178, USA.
Nat Biotechnol. 1996 Mar;14(3):339-42. doi: 10.1038/nbt0396-339.
Retrovirus-derived vectors are overwhelmingly preferred over other methods for ex vivo gene therapy because they provide permanent integration of foreign genes into cellular DNA. In comparison, cationic lipids mediate efficent gene transfer, but expression is transient. When we combined cationic lipids with retrovirus particles we obtained a significant enhancement of transduction efficiency, depending upon the type of lipid formulation and the dose used. The relative effectiveness of these cytofectins was: DOSPA:DOPE > DOTMA:DOPE > DOTAP, resulting in 60-, 37-, and 5-fold increases in transduction efficiency, respectively, at optimum dosage. The effect of polycationic DOSPA:DOPE was dependent upon the viral envelope glycoprotein, was attainable by lipid treatment of either cells or virus particles, was not enhanced by the addition of polybrene, and was inhibited by chloroquine. These results strongly suggested that DOSPA:DOPE act primarily by modulation of charge associated with the viral envelope and cell membrane, enhancing retroviral transduction, rather than by providing an alternative pathway of transfection. DOSPA:DOPE is useful for improving the efficiency of gene transfer as well as the sensitivity with which retroviruses can be detected in biological fluids.
在体外基因治疗中，逆转录病毒衍生载体比其他方法更受青睐，因为它们能将外源基因永久整合到细胞DNA中。相比之下，阳离子脂质介导高效的基因转移，但表达是短暂的。当我们将阳离子脂质与逆转录病毒颗粒结合时，根据脂质制剂的类型和使用剂量，转导效率得到了显著提高。这些细胞转染试剂的相对有效性为：DOSPA:DOPE > DOTMA:DOPE > DOTAP，在最佳剂量下，转导效率分别提高了60倍、37倍和5倍。聚阳离子DOSPA:DOPE的作用取决于病毒包膜糖蛋白，通过对细胞或病毒颗粒进行脂质处理均可实现，添加多聚赖氨酸不会增强其效果，而氯喹会抑制其作用。这些结果强烈表明，DOSPA:DOPE主要通过调节与病毒包膜和细胞膜相关的电荷来增强逆转录病毒转导，而不是通过提供另一种转染途径。DOSPA:DOPE有助于提高基因转移效率以及在生物流体中检测逆转录病毒的灵敏度。
