利用假病毒技术研究抗HIV-1药物的作用机制

The action mechanisms of anti-HIV-1 compounds with pseudovirus system

摘要:

采用假病毒系统对5种具有抗HIV-1活性的天然化合物的作用机制进行研究, 建立了一种可在普通实验室进行抗HIV-1作用机理研究的实验平台。通过构建假病毒系统, 利用定量PCR实验分析感染细胞内病毒生命周期早期的特异性DNA产物, 发现5种化合物可通过不同的作用机制在早期抑制HIV-1复制。部分化合物表现出新的作用机制, 如LC-1、LC-2可抑制HIV-1的入核。通过分析药物对2种不同的逆转录病毒(HIV-1和MLV)的感染性, 发现LC-1可特异地阻断HIV-1的复制, 而对MLV无影响。同时为了检测药物是否也会在病毒的生命周期晚期有作用,利用直接转染前病毒质粒的方式进行了验证。利用以上方法, 初步确定了这几种药物的作用靶点, 构建了一个有效的抗HIV-1药物作用机理的研究平台。

Abstract:

In this study, a pseudovirus system was constructed to investigate the anti-HIV mechanisms of five natural compounds. Three plasmids containing backbone, gag-pol of HIV-1 and VSVG were transfected to 293T cells to produce pseudotyped HIV-1 virus. Real-time PCR assay was used to explore the targets of these compounds by testing the specific viral products in the early life cycle of HIV-1. With this system, the targets of four compounds have been initially identified. LC-1 and LC-2 could block the nuclear import of HIV-1preintegration complex (PIC) into the nucleus. LC-3 could inhibit viral DNA integration to host genome. LC-4 could inhibit the activities of reverse transcriptase. Compounds LC1 and LC2 showed different action target from the existing drugs. Further study has been carried to explore the new potential action mechanism. This study showed that the pseudovirus system we constructed can be used as an assay platform to screen anti-HIV drugs.