NF Kappa Beta and the Regulation of HIV Latency

根除艾滋病毒和治愈感染者失败的主要原因是艾滋病毒建立了一种潜在的感染形式 在静息的CD4记忆T细胞中。虽然总共只有106-107个潜伏感染的细胞可能存在于感染的 患者，这些细胞的长期存活特性(44个月的T1/2)预测-70年的抗病毒治疗将是 消融这个蓄水池所需的能量。显然需要一种不同的策略来解决这个临床问题。一 该方法可能涉及通过刺激HIV LTR的转录激活来从这些细胞中清除病毒。 我们推测，核因子-B/Rel转录因子家族作为一种生理拮抗剂发挥着关键作用。 艾滋病毒潜伏期。我们建议进行一系列全面的研究，探索艾滋病毒潜伏期和 核因子-CB/Rel蛋白的作用。这些研究将开发一种新的HIV潜伏期细胞模型，J-Lat细胞，Bona 来自感染患者的真正潜伏感染的CD4记忆T细胞，以及慢病毒潜伏的灵长类动物模型。这些研究 将与Eric Verdin博士(项目1)、Matija Peterlin博士(项目3)和Tom博士进行密切和积极的合作 诺斯和保罗·卢西奥(B核心)共同组成了我们的PPG球队。在具体目标1中，我们将确定 转录因子NF-KB/Rel家族的成员是潜伏的HIV和Will最有效的激活剂 探索是否可以利用对核因子-KB/REL作用调节的新见解来逆转HIV潜伏期。在……里面 具体目标2，我们将使用染色质免疫沉淀(ChIP)分析来评估病毒潜伏期的分子基础 确定在病毒潜伏期和病毒潜伏期相反的条件下与HIV LTRKB增强子结合的细胞因子 活跃的病毒复制。在具体目标3中，我们将重点讨论核因子-kB与PTEFb之间有趣的相互作用。 被HIV Tat劫持的酶复合体，促进病毒基因的高水平表达。这些研究将剖析 核因子-KB与PTEFb组装的分子基础以及TAT的初始生产是否依赖于此 核因子-kB的作用。在具体目标4中，我们将推进对艾滋病毒潜伏期及其核因子-kB潜在拮抗作用的研究 进入活体模型系统。我们将调查_CB增强子的缺失是否在SHV病毒的HIV LTR中 增强恒河猴记忆CD4T细胞的潜伏感染，反之，是否替代具有构成活性的T细胞 细胞增强剂削弱潜伏期。最后，我们将探索已知的激活和维持核核因子-KB的制剂 前列环素和苯丁酸酯可减少潜伏感染这些SHIV的细胞的频率。这些研究 承诺促进我们对艾滋病毒潜伏期的分子、生化和细胞基础的理解，这反过来又会 可能导致新的理性方法来解决这一棘手的临床问题。

The eradication of HIV and cure of infected patients has failed chiefly because HIV establishes a latent form of infection in resting CD4 memory T-cells. Although a total of only 106-107 latently infected ceils may be present in infected patients, the long lived nature of these cells (T1/2 of 44 months) predicts that -70 years of antiviral therapy will be required for ablation of this reservoir. A different strategy is clearly required to attack this clinical problem. One approach could involve purging of the virus from these cells by stimulating transcriptional activation of the HIV LTR. We hypothesize that the NF-_B/Rel family of transcription factors plays a pivotal role as physiological antagonists of HIV latency. We propose to conduct a comprehensive set of studies exploring the molecular basis for HIV latency and the role played by NF-_cB/Rel proteins. These studies will exploit a new cellular model of HIV latency, J-Lat cells, bona fide latently infected CD4 memory T ceils from infected patients, and a primate model of lentiviral latency. These studies will involve close and active collaborations with Dr. Eric Verdin (Project 1) Dr. Matija Peterlin (Project 3) and Drs. Tom North and Paul Luciw (Core B) which together comprise our PPG team. In Specific Aim 1, we will determine which members of the NF-KB/Rel family of transcription factors function as the most potent activators of latent HIV and will explore whether new insights into the regulation of NF-KB/Rel action can be exploited to reverse HIV latency. In Specific Aim 2, we will assess the molecular basis of viral latency using chromatin immunoprecipitation (CHIP) assays to identify cellular factors that bind to the HIV LTR KB enhancers under the opposing conditions of viral latency and active viral replication. In Specific Aim 3, we will focus on the intriguing interplay of NF-KB with PTEFb, the cellular enzyme complex hijacked by HIV Tat to promote high level viral gene expression. These studies will dissect the molecular basis for NF-KB assembly with PTEFb and explore whether the initial production of Tat depends on this action of NF-KB. In Specific Aim 4, we will advance our studies of HIV latency and its potential antagonism by NF-KB into an in vivo model system. We will investigate whether deletion of the _cB enhancers in the HIV LTR of SHIV viruses enhances latent infection of rhesus memory CD4 T-cells and conversely whether substitution of a constitutively active T cell enhancer impairs latency. Finally, we will explore whether agents known to activate and sustain nuclear NF-KB action (prostratin and phenylbutyrate) decrease the frequency of cells latently infected with these SHIVs. These studies promise to advance our understanding of the molecular, biochemical, and cellular basis for HIV latency, which in turn may lead to new rational approaches to solving this difficult clinical problem.