Center for Proteomics and Bioinformatics, School of Medicine, Case Western Reserve University, 919 BRB 10900 Euclid Ave, Cleveland, OH 44106, USA.
J Virol Methods. 2014 Jan;195:164-9. doi: 10.1016/j.jviromet.2013.08.029. Epub 2013 Sep 8.
Fusion between the viral membrane of human immunodeficiency virus (HIV) and the host cell marks the end of the HIV entry process and the beginning of a series of post-entry events including uncoating, reverse transcription, integration, and viral gene expression. The efficiency of post-entry events can be modulated by cellular factors including viral restriction factors and can lead to several distinct outcomes: productive, latent, or abortive infection. Understanding host and viral proteins impacting post-entry event efficiency and viral outcome is critical for strategies to reduce HIV infectivity and to optimize transduction of HIV-based gene therapy vectors. Here, we report a combination reporter virus system measuring both membrane fusion and viral promoter-driven gene expression. This system enables precise determination of unstimulated primary CD4+ T cell subsets targeted by HIV, the efficiency of post-entry viral events, and viral outcome and is compatible with high-throughput screening and cell-sorting methods.
HIV 病毒的包膜与宿主细胞融合标志着 HIV 进入过程的结束和一系列后续事件的开始，包括脱壳、逆转录、整合和病毒基因表达。后续事件的效率可以被包括病毒限制因子在内的细胞因子调节，并导致几种不同的结果：有感染性的、潜伏的或无感染性的感染。了解宿主和病毒蛋白对后续事件效率和病毒结果的影响，对于减少 HIV 感染性和优化 HIV 为基础的基因治疗载体转导的策略至关重要。在这里，我们报告了一种组合报告病毒系统，该系统可测量膜融合和病毒启动子驱动的基因表达。该系统能够精确地确定 HIV 靶向的未受刺激的原发性 CD4+T 细胞亚群，后续病毒事件的效率以及病毒结果，并且与高通量筛选和细胞分选方法兼容。
