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.