Kwon Young Jik, Peng Ching-An
Department of Chemical Engineering, University of Southern California, Los Angeles, California 92697, USA.
Tissue Eng Part A. 2008 Sep;14(9):1497-506. doi: 10.1089/ten.tea.2007.0436.
Retroviral vectors are powerful tools for gene therapy and stem cell engineering. To improve efficiency of retroviral gene delivery, quantitative understanding of interactions of a retroviral vector and a cell is crucial. Effects of nonspecific adsorption of retrovirus on a cell via proteoglycans and receptor-mediated binding of retrovirus to a cell on overall transduction efficiency were quantified by combining a mathematical model and experimental data. Results represented by transduction rate constant, a lumped parameter of overall transduction efficiency, delineated that chondroitin sulfate C (CSC) plays dual roles as either enhancer or inhibitor of retroviral transduction, depending on its concentrations in the retroviral supernatant. At the concentration of 20 microg/mL, CSC enhanced the transduction efficiency up to threefold but inhibited more than sevenfold at the concentration of 100 microg/mL. Transduction rate constants for amphotropic retroviral infection of NIH 3T3 cells under phosphate-depleted culture condition showed a proportional relationship between cellular receptor density on a cell and transduction efficiency. It was finally shown that amphotropic retrovirus transduced human fibroblast HT1080 cells more efficiently than NIH 3T3 cells. On the contrary, the transduction efficiency of NIH 3T3 cells by vesicular stomatitis virus G protein pseudotyped retroviruses was eightfold higher than that of HT1080 cells. This study implies usefulness of using quantitative analysis of retroviral transduction in understanding and optimizing retroviral gene delivery systems for therapeutic approaches to tissue engineering.
逆转录病毒载体是基因治疗和干细胞工程的强大工具。为提高逆转录病毒基因递送效率，定量了解逆转录病毒载体与细胞间的相互作用至关重要。通过结合数学模型和实验数据，对逆转录病毒经蛋白聚糖非特异性吸附于细胞以及逆转录病毒与细胞受体介导的结合对整体转导效率的影响进行了量化。以转导率常数（整体转导效率的一个集总参数）表示的结果表明，硫酸软骨素C（CSC）根据其在逆转录病毒上清液中的浓度，对逆转录病毒转导起着增强剂或抑制剂的双重作用。在浓度为20μg/mL时，CSC将转导效率提高了三倍，但在浓度为100μg/mL时抑制作用超过七倍。在磷酸盐缺乏的培养条件下，嗜性逆转录病毒感染NIH 3T3细胞的转导率常数显示，细胞上的细胞受体密度与转导效率之间存在比例关系。最终表明，嗜性逆转录病毒对人成纤维细胞HT1080的转导效率高于NIH 3T3细胞。相反，水泡性口炎病毒G蛋白假型逆转录病毒对NIH 3T3细胞的转导效率比HT1080细胞高八倍。本研究表明，利用逆转录病毒转导的定量分析有助于理解和优化用于组织工程治疗方法的逆转录病毒基因递送系统。
