Functional genomic analysis of the CD8 T cell response to HIV vaccines in

描述(由申请人提供)：开发有效的艾滋病毒疫苗是全球最大的卫生挑战之一。然而，尽管进行了数十年的研究，还没有一种疫苗能够诱导对艾滋病毒的保护性免疫。最近，由于T细胞免疫和HIV病毒控制之间的关联，开发抗HIV疫苗的努力集中在优化T细胞对疫苗的反应上。然而，CD8T细胞对疫苗的最佳反应的分子相关性仍不清楚。这项研究将利用功能基因组学、细胞免疫学和动物模型来鉴定疫苗接种后CD8 T细胞最佳功能的分子特征。 在目标1中，我们将生成目前临床开发中的三种HIV疫苗诱导的四聚体分选HIV特异性CD8 T细胞的全基因组表达谱。我们将把这些HIV特异的CD8谱与抗原特异的CD8 T细胞进行比较，这些CD8 T细胞对针对痘苗病毒(VV)的疫苗产生反应--VV是有史以来最成功的疫苗之一。我们将确定每种疫苗特有的基因模式，以及诱导高功能CD8T细胞的疫苗(如VV)的共同特征。通过详细的计算分析，将确定赋予每个疫苗诱导的CD8 T细胞反应不同功能属性的候选机制。这将是第一次对疫苗诱导的抗原特异性CD8T细胞进行全面的转录分析，并将确定高功能CD8T细胞的新的分子相关性。 越来越多的数据表明，腺病毒(Ad5)载体疫苗可以诱导功能有限的CD8T细胞。在目标2中，我们将检验这一假设，即CD8T细胞对Ad5载体免疫原的反应上调的基因有助于它们的次优功能。基于我们在小鼠模型上的研究，并补充来自Aim 1的新数据，我们将使用功能基因组方法评估多达96个候选调节基因在Ad5/HIV诱导的CD8 T细胞中增加的作用。一系列高度平行的过表达研究和RNAi介导的基因敲除实验将验证候选基因作为抑制分子在人CD8T细胞和Ad5疫苗接种的小鼠模型中的作用。拟议中的研究将通过发现最佳疫苗诱导的T细胞免疫的新相关性，并确定疫苗接种后控制CD8 T细胞功能的机制，来加速艾滋病毒疫苗的开发。 与公共卫生相关：尽管进行了几十年的研究，但还没有任何疫苗能够产生强有力地预防艾滋病毒的免疫反应，我们仍然不知道保护性疫苗应该引发什么类型的免疫反应。这项提议将使用尖端技术：1)调查数千个免疫细胞对艾滋病毒疫苗做出反应的基因；2)确定表征最具功能的疫苗的基因模式；3)测试这些基因中哪些影响免疫系统的功能。这项研究的信息将有助于确定新的测试，以评估疫苗的工作效果，并发现对艾滋病毒疫苗做出更强免疫反应的新方法。

DESCRIPTION (provided by applicant): The development of an effective HIV vaccine is one of the greatest global health challenges. However despite decades of research, no vaccine has yet been able to elicit protective immunity to HIV. Recently, efforts to develop vaccines against HIV have focused on optimizing the T cell response to vaccination because of the association between T cell immunity and viral control in HIV. However, the molecular correlates of an optimal CD8 T cell response to vaccination remain unclear. This investigation will identify the molecular characteristics of optimal CD8 T cell function after vaccination, using functional genomics, cellular immunology and animal models. In Aim 1, we will generate genome-wide expression profiles of tetramer-sorted HIV-specific CD8 T cells elicited by three HIV vaccines in current clinical development. We will compare these HIV-specific CD8 profiles to antigen-specific CD8 T cells responding to vaccines against vaccinia virus (VV) - one of the most successful vaccines ever developed. We will identify patterns of genes that are unique to each type of vaccine and signatures that are common to those vaccines, such VV, that induce highly functional CD8 T cells. Candidate mechanisms that confer different functional attributes to each vaccine-induced CD8 T cell response will be identified using detailed computational analyses. This will be the first comprehensive transcriptional analysis of vaccine-induced antigen-specific CD8 T cells, and will identify new molecular correlates of highly functional CD8 T cells. Increasing data suggest that adenovirus (Ad5) vectored vaccines elicit CD8 T cells with limited functionality. In Aim 2 we will test the hypothesis that genes upregulated in CD8 T cells responding to Ad5-vectored immunogens contribute to their sub-optimal function. Based on our studies in a mouse model, and supplemented with new data from Aim 1, we will evaluate the role of up to 96 candidate regulator genes that are increased in Ad5/HIV-induced CD8 T cells using functional genomic approaches. A series of highly parallel over-expression studies and RNAi-mediated knockdown experiments will validate the role of candidate genes as inhibitory molecules in human CD8 T cells and in a mouse model of Ad5 vaccination. The proposed studies will accelerate HIV vaccine development by discovering new correlates of optimal vaccine-induced T cell immunity, and identifying the mechanisms that govern CD8 T cell function after vaccination. PUBLIC HEALTH RELEVANCE: Despite decades of research, no vaccines have yet been able to generate an immune response that robustly protects against HIV, and we still don't know what type of immune response should be elicited by a protective vaccine. This proposal will use cutting-edge technologies to: 1) survey thousands of genes expressed by immune cells responding to HIV vaccines; 2) identify the patterns of genes that characterize the most functional vaccines; and 3) test which of these genes influences the function of the immune system. Information from this study will help identify new tests to evaluate how well vaccines work, and discover new ways to make a stronger immune response to HIV vaccines.