Viral and Placental Determinants of HIV-1 Subtype C Mother-to-Child Transmission

HIV-1 C 亚型母婴传播的病毒和胎盘决定因素

• 批准号: 7223672
• 负责人: JESSE J KWIEK
• 金额: $ 9万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 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 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7223672
• 关键词: Acute; Address; Archives; Biological Assay; Biology; Biopsy; Blood; Breast Feeding; CCR5 gene; CXCR4 gene; Cells; Child; Cloning; Country; DNA Sequence Analysis; Data; Environment; Event; Genes; Genotype; Grant; HIV; HIV Core Protein p24; HIV Envelope Protein gp120; HIV Infections; HIV-1; Human; Immunoglobulin Variable Region; Immunohistochemistry; In Situ Hybridization; Incidence; Infant; Infection; Intervention; Length; Measures; Mediating; Mentors; Methods; Mothers; Nevirapine; Patient currently pregnant; Peripheral; Phase; Phenotype; Placenta; Plasma; Population; Pregnancy; Prophylactic treatment; Public Health; Rate; Reporting; Research Personnel; Risk Factors; Sampling; Score; Site; Staining method; Stains; Standards of Weights and Measures; Techniques; Testing; Tissues; Training; Tropism; Umbilical Cord Blood; Variant; Vertical Disease Transmission; Viral; Virus; Woman; cohort; experience; glycosylation; in utero; intrapartum; macrophage; novel; peripheral blood; prevent; programs; receptor; receptor expression; research study; scale up; transmission process; trophoblast

DESCRIPTION (provided by applicant): This proposal aims to comprehensively describe host and virus biology associated with HIV-1 subtype C (HIV-1C) mother-to-child transmission (MTCT). We hypothesize that HIV-1 C MTCT is a non-stochastic event mediated by both placental and viral determinants. To address this hypothesis, we will: 1) genotype and phenotype vertically transmitted envelope (env) genes, 2) characterize envelope genes isolated from archived placental biopsies, 3) describe the placental environment during MTCT. All experiments will use previously collected samples from a well-characterized cohort of 744 HIV-1 C-infected pregnant Malawian women characterized as nontransmitting (NT), in utero (IU) mother-offspring pairs (MOPs) and intrapartum (IP) MOPs. In a preliminary study, we enumerated HIV-1 C V1 A/2 env diversity in 22 MOPs and found that infants had significantly fewer variants than their mothers and confirmed that HIV env diversity is restricted during MTCT. During the mentored portion of this grant (Specific Aim 1), I will test the hypothesis that restriction is facilitated by specific env genotypes. This will be accomplished by cloning full-length envelope genes to: a) compare V1/V2 loop lengths, b) enumerate putative glycosylation sites, c) pseudotype the cloned env genes against CCR5, CxCR4, CD4hi and CD4lo cells. As an independent investigator, I will focus on ID MTCT and test the hypothesis that the placenta is a unique HIV-1 C compartment. Using a heteroduplex tracking assay, I will compare HIV-1 C env genes from matched peripheral blood, placental blood, placental biopsies, and cord blood (Specific Aim 2). Placental specific variants will be characterized using the methods acquired during the mentored phase. In Specific Aim 3, transmitted and placental specific variants will be psuedotyped against primary placental trophoblasts and Hofbauer cells. Finally, in Specific Aim 4, placental biopsies will be analyzed for HIV-1 C, CCR5, CD4, and CXCR4 expression. The results of this project will provide a comprehensive description of the HIV-1 C envelope genes and the placental environment conducive to in utero HIV-1 C MTCT.

描述（由申请人提供）：该提案旨在全面描述与HIV-1亚型C（HIV-1C）母亲到孩子传播（MTCT）相关的宿主和病毒生物学。我们假设HIV-1 C MTCT是由胎盘和病毒决定因素介导的非传染性事件。为了解决这一假设，我们将：1）基因型和表型垂直传播的包膜（ENK）基因，2）表征从存档的胎盘活检中分离出来的包膜基因，3）描述MTCT期间的胎盘环境。所有实验将使用先前收集的744个HIV-1 C感染的马拉维孕妇的样品，该孕妇在子宫（IU）母亲春季（MOPS）和全部（IP）MOPS中，其特征为非翻译（NT）。在一项初步研究中，我们列举了22 MOPS中的HIV-1 C V1 A/2环境的多样性，发现婴儿的变体明显少于母亲，并证实HIV ENV在MTCT期间受到限制。在这笔赠款的指导部分（特定目标1）期间，我将检验以下假设：限制是由特定的ENV基因型促进的。这将通过将全长的包膜基因克隆到：a）比较V1/V2回路长度，b）列举推定的糖基化位点，c）对CCR5，CXCR4，CXCR4，CD4HI和CD4LO细胞进行伪装。作为一名独立研究者，我将重点关注ID MTCT，并检验胎盘是独特的HIV-1 C室的假设。使用杂化跟踪测定法，我将比较来自匹配的外周血，胎盘血，胎盘活检和脐带血的HIV-1 C ENV基因（特定的AIM 2）。将使用在指导阶段获得的方法来表征胎盘特定的变体。在特定的目标3中，将针对原代胎盘滋养细胞和霍夫鲍尔细胞进行传播和胎盘特异性变体。最后，在特定的目标4中，将对HIV-1 C，CCR5，CD4和CXCR4表达进行分析胎盘活检。该项目的结果将提供对HIV-1 C信封基因的全面描述以及在子宫内HIV-1 C MTCT中有利于胎盘环境。