HIV-1假病毒包装的重要影响因素分析

目的]分析影响HIV-1假病毒包装的主要因素,建立该假病毒包装的优化条件.方法]用单因素分析对比的方法,比较了对病毒包装效果有重要影响的转染试剂、转染试剂与质粒的数量、培养基血清类型、细胞周期阶段对于病毒包装效果的影响.结果]对45批150盘病毒包装结果分析显示,使用PEI转染试剂成本低,转染效果好,其N/P在8-40均可以产生高活性的包装病毒；用进口血清包装时,相对于国产血清结果重复性高；细胞周期处于S及G2/M期时转染质粒相对于S以及G0/G1期有较高的病毒活性.结论]用PEI为转染试剂可以包装出高活性的病毒,包装获取高活性病毒可以通过质粒与PEI梯度比例以及血清类型构成多个组合策略来实现.

Analysis of factors influencing the infectivity of HIV-1 pseudotyped virus during packaging by cell line

Objective] A serials of factors which may have influence on producing the HIV-1 pseudotyped virus are analysed to establish an optimal way to generate this type of virus. Methods] A set of factors such as the transfection reagents, the ratios of plasmid and transfection, the serum in the media and cell cycle is being analysis to find the optimal condition. Results] The optimal N/P is range from 8 to 40 , the serum can influence the stability of pseudotype virus and in the stage S and G2/M, transfection efficiency is high than the stage G0/G1. Conclusion] High titers of pseudotyped HIV-1 virus can be obtained by means of using transfection reagents PEI and through combinatorial optimization of PEI, plasmid amount and FBS type.