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

描述(由申请人提供)：本R03申请旨在提供数据，支持由药物逃逸突变产生的HIV-1蛋白酶新表位的CD4T细胞反应有助于控制耐药变异的生长这一基本假设。经PI治疗的HIV-1感染者(HIV-1)中有相当大比例的患者发生耐药突变，CD8 T细胞对这种药物诱导的突变表位的反应已有文献记载。已有研究表明，在接受抗逆转录病毒(ARV)治疗的患者中，药物压力下针对表位的这种T细胞反应可能会使突变株处于免疫控制之下，或延迟耐药突变的出现。鉴于CD4T细胞反应在维持CD8T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CD4T细胞对PI诱导突变的位置的识别。该研究将有一个横截面(目标1)和一个纵向臂(目标2)。为了寻找针对突变的蛋白水解酶表位(AIM 1)的CD4T细胞反应，我们将合成代表野生型和新表位序列的多肽，包含PI诱导的主要突变位点，并预测在电子计算机上与多个HLA-DR分子结合。为了评估它们的混杂，我们将评估它们在体外结合多个HLA-DR分子的能力。随后，我们将通过干扰素-γELISPOT、CFSE增殖和流式细胞术检测PI处理的HIV-1患者的T细胞对这些多肽的反应。识别的多肽序列将与内源性HIV-1蛋白酶序列进行比较。在纵向臂(目标2)中，我们将研究其HIV-1分离株在1年观察期内发生新的蛋白酶基因突变的患者。在这些患者中，我们将比较对新表位产生或失败的CD4T细胞反应的组之间的疾病演变。我们希望获得关于药物和免疫压力的融合是否会影响导致免疫逃逸或耐药性的突变的信息。将获得的信息可能有助于开发免疫策略，这些策略可用于增强对突变HIV-1的免疫反应，并对抗耐PI的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.