Improved retroviral transduction of hematopoietic progenitors by combining methods to enhance virus-cell interaction.
Improved retroviral transduction of hematopoietic progenitors by combining methods to enhance virus-cell interaction.
复制标题
通过结合增强病毒与细胞相互作用的方法来改善造血祖细胞的逆转录病毒转导。
DOI：
10.1038/sj.leu.2401672
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发表时间：
2000
期刊：
影响因子：
11.4
通讯作者：
Verfaillie,CM
中科院分区：
文献类型：
--
作者：
Liu,H;Hung,Y;Wissink,SD;Verfaillie,CM
成功进行逆转录病毒转导所需的因素之一是病毒颗粒与靶细胞之间的接触。我们假设，通过不同机制改善病毒与靶细胞相互作用的药物组合将提高转导效率。我们通过半固体培养计数G418抗性克隆，检测了两性逆转录病毒载体G1Na对白血病K562细胞、原代正常和慢性粒细胞白血病CD34+细胞的转导效率。我们评估了重组纤维连接蛋白片段CH296、阳离子脂类或跨孔流系统单独或联合应用改善逆转录病毒转导的能力。脂类或CH296可使K562细胞的转导能力提高1.5~2倍，而两者联合使用则可使转导能力提高2.5倍。在跨井流过系统中转导K562细胞的转导能力提高了三倍。正常(NL)CD34+CFCs的转导在脂质作用下提高了10倍，在CH296作用下提高了20倍。脂质和CH296具有协同作用。跨井通流系统可将正常CD34+CFCs的转导能力提高30倍。最后，与对K562细胞的研究类似，CH296或脂类对CML CFC的转导提高了两到三倍，而这两种药物的组合具有协同作用。我们的结论是，任何加强病毒颗粒和靶细胞之间接触的物理手段都会改善转导。两种具有不同作用机制的方法对转导有相加或协同作用。
One of the factors required for successful retroviral transduction is contact between viral particles and target cells. We hypothesized that combining agents that improve virus–target cell interaction via different mechanisms will increase transduction efficiency. We examined the transduction efficiency of leukemic K562 cells, primary normal and chronic myelogenous leukemia CD34+ cells with the amphotropic retroviral vector, G1Na, packaged in PA317 by enumerating G418-resistant colonies in semisolid media. We evaluated the ability of the recombinant fibronectin fragment, CH296, cationic lipids, or a transwell flow-through system, alone or in combination to improve retroviral transduction. Transduction of K562 cells improved 1.5 to two-fold with lipids or CH296, while their combination improved transduction 2.5-fold. Transduction of K562 cells in the transwell flow-through system improved transduction three-fold. Transduction of normal (NL) CD34+ CFC improved 10-fold with lipids and 20-fold with CH296. Lipid and CH296 had synergistic effects. The transwell flow-through system improved transduction of normal CD34+ CFC 30-fold. Finally, similar to what was seen for K562 cells, transduction of CML CFC improved two-to three-fold with either CH296 or lipids, whereas the combination had synergistic effects. We conclude that any physical means that enhances contact between viral particles and target cells improves transduction. Two such methods that have different action mechanisms have additive or synergistic effects on transduction.