Elucidation of the Mechanisms of CD8+ T Cell Noncytolytic Antiviral Response

描述（由申请人提供）：感染细胞。这种类型的非细胞溶解性抑制称为CD8+ T细胞非细胞溶解性抗病毒反应（CNAR）。CNAR的机制尚未明确，但其重要性已被CNAR与hiv感染者健康临床状态的相关性所证明。我们的长期目标是了解CNAR的机制，这样我们就可以学习如何引发或模仿这种反应。目前的研究表明，一种未知的分子或分子复合体在病毒转录开始时阻断了HIV-1的复制。我们的目标是分析转录调控的两个分支-宿主表观遗传学和病毒遗传元件-以确定CNAR的机制和关键分子。建议的工作将集中在CNAR诱导转录抑制因子如组蛋白去乙酰化酶来抑制HIV-1前病毒的转录的假设上；并且HIV-1可以通过突变其基因组来消除这些转录抑制因子的作用。初步数据表明，抑制组蛋白去乙酰化可导致CNAR降低；并且随着时间的推移，遗传相关的HIV-1分离株对CNAR的敏感性会发生变化。我们将通过在敏感病毒和耐药病毒之间创造嵌合病毒来确定赋予耐药性的遗传因素。此外，我们将使用染色质免疫沉淀来检测CNAR抑制培养中CD4+ T细胞HIV-1长末端重复中组蛋白去乙酰化酶1的募集及其对核心组蛋白乙酰化的影响。我们的研究将更好地了解介导CNAR的分子，CNAR的机制，并有助于开发模拟或诱导这种免疫反应的新型抗病毒药物。拟议的研究将确定CD8+ T细胞如何在不杀死感染细胞的情况下抑制HIV-1。确定这种免疫反应的机制将提供对HIV感染中宿主-病原体相互作用的更好理解，并可能导致新的HIV治疗方法的发展。

DESCRIPTION (provided by applicant): the infected cell. This type of noncytolytic suppression is called CD8+ T cell noncytolytic antiviral response (CNAR). The mechanisms of CNAR are as yet uncharacterized, but its importance is evidenced by CNAR's correlation with healthy clinical status in HIV-infected individuals. It has been a long-term goal of ours to discern the mechanisms of CNAR, so that we can learn how to elicit or mimic this response. The current research suggests that an unknown molecule or complex of molecules block HIV-1 replication at the initiation of virus transcription. We aim to analyze two branches of transcription regulation-host epigenetics and viral genetic elements-to identify the mechanisms of CNAR and the key molecules responsible. The proposed work will be focused on the hypothesis that CNAR induces transcriptional repressors such as histone deacetylases to suppress transcription from HIV-1 provirus; and that HIV-1 can mutate its genome to negate the effects of these transcriptional repressers. Preliminary data has shown that inhibiting histone deacetylation results in a decrease in CNAR; and that genetically-related HIV-1 isolates evolve differing sensitivities to CNAR over time. We will identify genetic elements that confer resistance by creating chimeric viruses between sensitive and resistant viruses. Also, we will use chromatin immunoprecipitation to examine the recruitment of histone deacetylase 1 and its effects on acetylation of core histones within the HIV-1 long terminal repeat of CD4+ T cells in CNAR suppression cultures. Our study will provide a better understanding of the molecules that mediate CNAR, the mechanisms of CNAR, and aid in developing novel antivirals that mimic or induce this immune response. The proposed research will define how CD8+ T cells suppress HIV-1 without killing the infected cell. Determining the mechanism of this immune response will provide a greater understanding of the host pathogen-interaction in HIV infection and potentially lead to the development of novel HIV therapeutics.