The Release of HIV-1 is not Impacted by Levels of Tetherin in Human Hela Cell Lineage

M. S. Cunha, L. J. Costa
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Abstract

Tetherin is an interferon-induced transmembrane protein that maintains enveloped viruses tethered to cell membranes. Several viruses have developed proteins capable to counteract tetherin from their respective hosts, e.g. accessory protein Vpu from Human Immunodeficiency Virus (HIV-1) and Nef from some Simian Imunodeficiency Viruses (SIVs). Studies involving the role of tetherin in the release of viruses were done in the context of tetherin superexpression. The real impact of tetherin in a physiological condition is still a matter of debate. Therefore, our group started to investigate the differences in levels of tetherin and in the release of HIV-1 and SIVcpz in HeLa cells, a cell line that constitutively express tetherin. HeLa cells were transfected with HIV-1 WT, HIV-1 ΔNef and HIV-1 ΔVpu infectious clones and after 48 hours, cell lysates were collected and submitted to SDS-PAGE to assess tetherin levels and the levels of released viruses were analyzed by ELISA αp24. As expected, we observed about 25% reduction on levels of tetherin from both HIV-1 WT and HIV-1ΔNef when compared with HIV-1 ΔVpu. However, the levels of released virus were not altered despite of the down-modulation of tetherin. Our results show that endogenous levels of human tetherin are not detrimental for release of HIV-1.
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人类Hela细胞系中Tetherin水平不影响HIV-1的释放
系带蛋白是一种干扰素诱导的跨膜蛋白,可维持被包膜病毒系在细胞膜上。一些病毒已经开发出能够抵抗来自各自宿主的链链蛋白的蛋白质,例如来自人类免疫缺陷病毒(HIV-1)的辅助蛋白Vpu和来自某些猿猴免疫缺陷病毒(siv)的辅助蛋白Nef。涉及tetherin在病毒释放中的作用的研究是在tetherin超表达的背景下进行的。tetherin在生理条件下的真正影响仍然是一个有争议的问题。因此,我们的研究小组开始研究在HeLa细胞(一种组成性表达tetherin的细胞系)中tetherin水平以及HIV-1和SIVcpz释放的差异。用HIV-1 WT、HIV-1 ΔNef和HIV-1 ΔVpu感染克隆转染HeLa细胞,48h后收集细胞裂解液,提交SDS-PAGE检测tetherin水平,ELISA αp24检测释放的病毒水平。正如预期的那样,我们观察到与HIV-1 ΔVpu相比,HIV-1 WT和HIV-1ΔNef的tetherin水平降低了约25%。然而,尽管tetherin下调,但释放的病毒水平并未改变。我们的研究结果表明,内源性的人tetherin水平对HIV-1的释放没有危害。
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