HIV-1假病毒包装的重要影响因素分析

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

摘要:

【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。

Abstract:

[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.