Zhang H, Zhang Y, Spicer T P, Abbott L Z, Abbott M, Poiesz B J
Department of Microbiology/Immunology, SUNY Health Science Center at Syracuse, New York 13210.
AIDS Res Hum Retroviruses. 1993 Dec;9(12):1287-96. doi: 10.1089/aid.1993.9.1287.
Extracellular HIV-1 virions purified from cell culture supernatants have been found to contain viral DNA that is the result of partial reverse transcription within the virus particles. Our data supported these observations and further indicated that the ratio of genomic RNA to viral DNA was approximately 10(3):1 for the "strong stop" (R-U5) region and 10(5):1 for the gag region. We have shown that, in the absence of detergent, large amounts of DNase-resistant viral DNA can be synthesized within intact HIV-1 virions, indicating that this phenomenon is not dependent on perturbation of the viral envelope. Nascent viral DNA synthesis also occurred in purified virions incubated at 37 degrees C in cell-free human physiological fluids including seminal plasma, blood plasma, breast milk, and fecal fluid. In vitro HIV-1 infection assays, in which HIV-1 DNA synthesis was initiated in HIV-1 virions by prior incubation with deoxyribonucleoside triphosphates, demonstrated that virus particles so treated had an increased infectious titer over untreated virions when incubated with target human T cells. Our data suggest that HIV-1 virion-associated DNA synthesis may occur in vivo and may impact on the efficiency of intra- and interhost virus transmission. If so, this phenomenon should prove to be an important target for antiviral therapeutic strategies.
从细胞培养上清液中纯化出的细胞外HIV-1病毒粒子被发现含有病毒DNA，这是病毒粒子内部分逆转录的结果。我们的数据支持了这些观察结果，并进一步表明，对于“强终止”（R-U5）区域，基因组RNA与病毒DNA的比例约为10³:1，而对于gag区域则为10⁵:1。我们已经表明，在没有去污剂的情况下，完整的HIV-1病毒粒子内可以合成大量抗DNase的病毒DNA，这表明这种现象不依赖于病毒包膜的扰动。在包括精液、血浆、母乳和粪便液在内的无细胞人类生理液体中于37℃孵育的纯化病毒粒子中也发生了新生病毒DNA的合成。在体外HIV-1感染试验中，通过事先与脱氧核糖核苷三磷酸孵育在HIV-1病毒粒子中启动HIV-1 DNA合成，结果表明，当与靶人类T细胞孵育时，经过如此处理的病毒粒子比未处理的病毒粒子具有更高的感染滴度。我们的数据表明，HIV-1病毒粒子相关的DNA合成可能在体内发生，并可能影响宿主内和宿主间病毒传播的效率。如果是这样，这一现象应该被证明是抗病毒治疗策略的一个重要靶点。
