HIV-1 PROTEASE CD4+ T CELL EPITOPES AND DRUG-INDUCED MUTATIONS

描述（由申请人提供）：这项R03申请旨在提供数据来支持潜在的假设，即CD4+ T细胞对药物逃逸突变产生的HIV-1蛋白酶新表位的反应有助于控制耐药变异的生长。经pi治疗的HIV-1感染（HIV-1+）患者中有相当大比例出现耐药性突变，并且CD8+ T细胞对这种药物诱导的突变表位的反应已被证实。已有研究表明，在接受抗逆转录病毒（ARV）治疗的患者中，这种针对药物压力的T细胞应答可能使突变体处于免疫控制之下，或延迟耐药突变的出现。鉴于CD4+ T细胞反应在维持CD8+ T细胞记忆中的重要性，我们将首次研究pi治疗的HIV-1 +患者的CD4+ T细胞对pi诱导突变位点的识别。该研究将包括横断面研究（目标1）和纵向研究（目标2）。为了寻找CD4+ T细胞对突变的蛋白酶表位的反应（Aim 1），我们将合成代表野生型和新表位序列的肽，包含主要的pi诱导突变位点，并预测在硅中与多个HLA-DR分子结合。为了评估它们的混杂性，我们将评估它们在体外结合多种HLA-DR分子的能力。随后，我们将通过ifn - γ ELISPOT、CFSE增殖和流式细胞术检测pi治疗的HIV-1 +患者的T细胞对这些肽的反应。识别的肽序列将与内源性HIV-1蛋白酶序列进行比较。在纵向组（Aim 2）中，我们将研究在1年的观察期内HIV-1分离株出现蛋白酶基因新突变的患者。在这些患者中，我们将比较对新表位产生CD4+ T细胞反应或未产生CD4+ T细胞反应的组之间的疾病演变。我们期望获得关于药物和免疫压力的趋同是否影响导致免疫逃逸或耐药性的突变的信息。获得的信息可能有助于制定免疫策略，可用于增强对突变HIV-1的免疫反应，并对抗pi耐药HIV-1毒株的出现。

DESCRIPTION (provided by applicant): This R03 application aims at providing data in support of the underlying hypothesis that CD4+ T cell responses towards HIV-1 protease neoepitopes created by drug escape mutations can help to control the growth of resistant variants. A significant proportion of Pi-treated HIV-1 infected (HIV-1+) patients develop resistance mutations, and CD8+ T cell responses against such drug-induced mutant epitopes have been documented. It has been suggested that such a T cell response against epitopes under drug pressure may keep mutants under immunological control, or delay the emergence of drug resistance mutations, in patients undergoing antiretroviral (ARV) treatment. Given the importance of CD4+ T cell responses in maintaining CD8+ T cell memory, we will investigate, for the first time, the recognition of sites of Pi-induced mutations by CD4+ T cells of Pi-treated HIV-1 + patients. The study will have a cross-sectional (Aim 1) and a longitudinal arm (Aim 2). To search for CD4+ T cell responses against mutant protease epitopes (Aim 1), we will synthesize peptides representing wild-type and neoepitope sequences, incorporating major Pi-induced mutation sites and predicted to bind to multiple HLA-DR molecules in silico. To evaluate their promiscuity, we will assess their ability to bind multiple HLA-DR molecules in vitro. Subsequently, we will test the T cell responses from Pi-treated HIV-1 + patients to such peptides by IFN-gamma ELISPOT, CFSE proliferation and flow cytometry. Sequences of recognized peptides will be compared to endogenous HIV-1 protease sequence. In the longitudinal arm (Aim 2), we will study patients whose HIV-1 isolates developed new mutations in the protease gene along the 1-year observation period. Among such patients, we will compare disease evolution between the groups that developed or failed to develop CD4+ T cell responses to neoepitopes. We expect to obtain information as to whether the convergence of drug and immune pressure influences mutations leading to immune escape or drug resistance. The information to be obtained may help develop immune strategies that could be applied to enhance the immune response against mutant HIV-1 and fight the emergence of Pi-resistant HIV-1 strains.