SIV antisense epitopes as novel markers of latency

 描述：HIV建立潜伏感染，阻碍彻底治愈的努力。这种治愈的可能性取决于识别、量化并最终消除潜伏感染细胞的新策略。一个理想的方法是识别潜伏感染的细胞，并在不需要重新激活潜伏病毒的情况下消除它们。在这里，基于对HIV和SIV转录和翻译的新见解，我们假设存在一种新的潜伏感染细胞标记。具体地说，我们假设潜伏感染与水平升高有关。 病毒的反义转录和翻译。已发表的关于HIV的数据和我们对恒河猴SIV感染的初步数据表明，当细胞不那么活跃时，感染的CD4+T细胞中的反义转录更加突出，并且这些独特的转录本中的许多是多腺化的，可以被翻译。此外，我们已经确定了在SIV和HIV感染期间启动的T细胞反应，这些反应针对的是来自反义转录产物翻译的令人惊讶的保守多肽。为了验证我们的假设，在目标1中，我们将使用SIV感染的恒河猴和HIV-1感染的人类细胞比较潜伏感染的CD4T细胞和激活的和感染的CD4T细胞之间的病毒转录(正义和反义)。为了做到这一点，我们将使用一种新的链特定的深度测序方法。目标1的结果将把我们的努力集中在目标2上。在目标2中，我们将使用识别反义编码表位的T细胞，并测量它们杀死潜伏感染与激活和感染的CD4T细胞的能力。总之，这项提议有可能识别一种新的潜伏感染细胞的标记，并提出一种消除它们的方法。这些数据可能被证明对开发艾滋病毒治疗方法的努力很重要。

 DESCRIPTION: HIV establishes latent infection, hampering efforts for a complete cure. The possibility of such a cure rests on new strategies to identify, quantify and ultimately eliminate latently infected cells. An ideal approach would be to identify latently infected cells and eliminae them without the need to reactivate the latent virus. Here, we posit the existence of a novel marker of latently infected cells, based on new insights into HIV and SIV transcription and translation. Specifically, we hypothesize that latent infection is associated with increased levels of viral antisense transcription and translation. Published data on HIV and our preliminary data with SIV infection of rhesus macaques indicate that antisense transcription in infected CD4+ T cells is more prominent when the cells are less activated, and that many of these unique transcripts are polyadenylated and could be translated. Further, we have identified T cell responses mounted during SIV and HIV infection that target surprisingly conserved peptides derived from translation of antisense transcripts. To test our hypothesis, in Aim 1, we will compare viral transcription (sense and antisense) between latently infected CD4 T cells and activated and infected CD4 T cells using both SIV-infected rhesus macaque and HIV-1 infected human cells. To do this, we will use a novel strand-specific deep sequencing approach. Results from Aim 1 will focus our efforts in Aim 2. In Aim 2, we will use T cells that recognize antisense-encoded epitopes and measure their ability to kill latently infected vs. activated and infected CD4 T cells. In sum, this proposal has the potential to identify a novel marker of latently infecte cells and suggest a means by which to eliminate them. These data could prove important to efforts to develop a cure for HIV.