Sakoda Tsuyoshi, Kasahara Nori, Kedes Larry, Ohyanagi Mitsumasa
Department of Internal Medicine, Division of Coronary Heart Disease;
Exp Clin Cardiol. 2007 Fall;12(3):133-8.
Lentivirus vectors provide a delivery system that can both transduce nondividing cells and integrate transgenes into the genome of target cells without cytotoxicity. However, their relatively low transduction efficiency presents a significant obstacle to progress.
In the present paper, a simple and easy method using calcium phosphate (CaPi) to enhance the efficiency of lentivirus gene transfer in both vascular smooth muscle cells and cardiac myocytes is reported.
Delivery of lentivirus vectors in the presence of CaPi coprecipitates increased vector-encoded transgene expression up to 13-fold. Of interest, the magnitudes of enhancement of transgene expression by CaPi coprecipitates in 293T cells, vascular smooth muscle cells and cardiac myocytes were greater during brief periods (10 min and 120 min) of virus-cell contact than during long periods (16 h). Moreover, with a short duration of incubation with CaPi coprecipitates (up to 120 min), there was little evidence of direct cell toxicity. CaPi coprecipitates had no effect on host range specificity of ecotropic viruses and thus appears to enhance transduction efficiency physiologically by facilitating physical interaction between virus and cell.
These data show that lentivirus with CaPi coprecipitates increases both the efficiency and the speed of gene transfer. These approaches provide an efficient method and an improved tool for research and possibly for therapy of cardiovascular diseases.
慢病毒载体提供了一种递送系统，它既能转导非分裂细胞，又能将转基因整合到靶细胞基因组中而无细胞毒性。然而，其相对较低的转导效率成为研究进展的重大障碍。
本文报道一种使用磷酸钙（CaPi）提高慢病毒基因在血管平滑肌细胞和心肌细胞中转移效率的简单易行方法。
在存在CaPi共沉淀物的情况下递送慢病毒载体，可使载体编码的转基因表达增加达13倍。有趣的是，在病毒与细胞短暂接触（10分钟和120分钟）期间，CaPi共沉淀物在293T细胞、血管平滑肌细胞和心肌细胞中增强转基因表达的幅度大于长期接触（16小时）期间。此外，与CaPi共沉淀物孵育时间较短（长达120分钟）时，几乎没有直接细胞毒性的证据。CaPi共沉淀物对嗜亲性病毒的宿主范围特异性没有影响，因此似乎通过促进病毒与细胞之间的物理相互作用在生理上提高转导效率。
这些数据表明，带有CaPi共沉淀物的慢病毒可提高基因转移的效率和速度。这些方法为心血管疾病的研究乃至治疗提供了一种有效的方法和改进的工具。
