Impact of HIV-1 Genotype on Therapy Response in Children

描述(申请人提供)：长期目标是检测艾滋病毒感染儿童和青少年的艾滋病毒演变和免疫功能，这些儿童和青少年在启动含有抗逆转录病毒疗法的蛋白酶抑制剂[ART]后，重建免疫[免疫成功，IS]，但未能控制病毒复制[病毒失败，VF]。研究将集中在一组独特的接受治疗的患者身上，这些患者尽管携带了可以预测疾病进展的病毒载量，但仍保持了免疫重建。这一结果组为病毒/宿主细胞相互作用的研究提供了一种新的范例，这可能导致新的治疗策略。这一提议背后的假设是，ART诱导的不协调反应是多因素的，涉及病毒的表型属性、胸腺的功能完整性以及正在进行的病毒复制对免疫的影响。为了确定VF/IS响应涉及的机制，提出了三个具体目标： 具体目的1.研究VF/IS患者外周血中GAG和蛋白水解酶[PR]氨基酸替换的进化动态。在ART期间在VF/IS个体中形成的GAG和PR氨基酸序列中遗传标记的积累和调节将用于评估：[A]血浆和CD4、CD45RO、T淋巴细胞和[B]中病毒的区隔。CD_4、CD_(45)RA T淋巴细胞中的病毒贮存库。 具体目的2.确定VF/IS个体GAG和PR中优先促进胸腺细胞限制性复制的遗传决定因素。带有来自不和谐患者的Gag/PR区域的重组病毒将被构建并在培养中进行评估：[A]比较治疗前后来自VF/IS患者的病毒的Gag/PR区域在胸腺细胞和PBMC体外复制的能力。绘制GAG和PR中有助于优先限制胸腺细胞复制能力的遗传决定因素图。 具体目标3.确定正在进行的病毒复制对病毒和免疫结果不一致的个体的免疫力的影响。具体研究将评估：[A]胸腺输出，[B.]胸腺T细胞活化和分化后，和[C.]VS/IS和VF/IS结局组对新抗原的功能性免疫反应。

DESCRIPTION (provided by applicant): The long-term objective is to examine HIV evolution and immune function in HIV-infected children and adolescents who, following initiation of protease inhibitor containing antiretroviral therapy [ART], reconstitute immunity [Immune success, IS] but fail to control viral replication [viral failure, VF]. Studies will focus on a unique cohort of treated patients who have sustained immune reconstitution in spite of viral loads that would predict disease progression. This outcome group presents a novel paradigm for the investigation of virus/host cell interactions that are likely to lead to novel new therapeutic strategies. The hypothesis underlying the proposal is that ART-induced discordant responses is multifactorial, involving phenotypic properties of the virus, functional integrity of the thymus, and the impact of ongoing viral replication on immunity. To determine the mechanisms involved in VF/IS responses, three specific aims are proposed: Specific Aim 1. To examine the evolutionary dynamics of amino acid substitutions in Gag and protease [PR] within the peripheral blood compartments of VF/IS individuals. Accumulation and modulation of genetic markers in Gag and PR amino acid sequences that develop in VF/IS individual during ART will be used to evaluate: [A.] compartmentalization of viruses in plasma and in CD4 CD45RO T lymphocytes and [B.] reservoirs of virus in CD4 CD45RA T lymphocytes. Specific Aim 2. To identify genetic determinants in gag and PR from VF/IS individuals that contribute preferentially to restricted replication in thymocytes. Recombinant viruses with gag/PR regions derived from discordant patients will be constructed and evaluated in culture to: [A.] compare pre- and post therapy gag/PR regions from viruses from VF/IS individuals with respect to their capacity to replicate in thymocytes and PBMC ex vivo and [B.] map genetic determinants in Gag and PR that contribute to preferential restriction of replicative capacity in thymocytes. Specific Aim 3. To determine the impact of ongoing viral replication on immunity among individuals who have discordant viral and immune outcomes. Specific studies will evaluate: [A.] thymic output, [B.] post thymic T cell activation and differentiation, and [C.] functional immune response to neoantigen between VS/IS and VF/IS outcome groups.