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

HIV-1 蛋白酶 CD4 T 细胞表位和药物诱导的突变

• 批准号: 7383145
• 负责人: EDECIO CUNHA-NETO
• 金额: $ 5.3万
• 依托单位: UNIVERSITY OF SAO PAULO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7383145
• 关键词: 5-(6)-carboxyfluorescein diacetate succinimidyl ester; Address; Affect; Alleles; Anti-Retroviral Agents; Binding; Biological Assay; Blood specimen; CD4 Positive T Lymphocytes; CD8B1 gene; Cell Count; Cells; Clinical assessments; Computer Simulation; Data; Disease; Drug resistance; Endopeptidases; Epitopes; Evaluation; Evolution; Exclusion Criteria; Flow Cytometry; Goals; Growth; HIV-1; HIV-1 protease; HLA-DR Antigens; Immune; Immune response; In Vitro; Induced Mutation; Interferon Type II; Measurement; Memory; Mononuclear; Mutation; Numbers; Patients; Peptide Hydrolases; Peptides; Pharmaceutical Preparations; Phenotype; Plasma; Production; Proliferating; Protease Gene; Protease Inhibitor; RNA; Resistance; Resistance development; Site; Surface; T memory cell; T-Lymphocyte; T-Lymphocyte Epitopes; Testing; Time; Upper arm; Variant; Viral; design; enzyme linked immunospot assay; fighting; mutant; peripheral blood; pressure; response

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.

描述（由申请人提供）：本R 03申请旨在提供数据支持以下基本假设：CD 4 + T细胞对药物逃逸突变产生的HIV-1蛋白酶新表位的应答有助于控制耐药变体的生长。相当大比例的PI治疗的HIV-1感染（HIV-1+）患者发生耐药突变，并且已经记录了针对这种药物诱导的突变表位的CD 8 + T细胞应答。有人认为，在药物压力下，这种针对表位的T细胞应答可能会使突变体处于免疫控制之下，或者在接受抗逆转录病毒（ARV）治疗的患者中延迟耐药性突变的出现。鉴于CD 4 + T细胞应答在维持CD 8 + 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CD 4 + T细胞对Pi诱导突变位点的识别。本研究将有一个横截面（目标1）和一个纵向臂（目标2）。为了寻找针对突变蛋白酶表位的CD 4 + T细胞应答（Aim 1），我们将合成代表野生型和新表位序列的肽，其并入主要的Pi诱导的突变位点，并预测在计算机模拟中结合多个HLA-DR分子。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增殖和流式细胞术测试来自Pi治疗的HIV-1 +患者的T细胞对这些肽的应答。将识别肽的序列与内源性HIV-1蛋白酶序列进行比较。在纵向臂（目标2）中，我们将研究HIV-1分离株在1年观察期内沿着蛋白酶基因出现新突变的患者。在这些患者中，我们将比较对新表位产生或未能产生CD 4 + T细胞应答的组之间的疾病演变。我们希望获得有关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。获得的信息可能有助于开发免疫策略，可用于增强对突变型HIV-1的免疫反应，并对抗Pi耐药HIV-1菌株的出现。