Analysis of human uterine mucosal cells as targets of HIV-1 infection

人类子宫粘膜细胞作为 HIV-1 感染靶点的分析

• 批准号: 8925576
• 负责人: JOHN Christopher KAPPES
• 金额: --
• 依托单位: BIRMINGHAM VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-10-01 至 2018-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8925576
• 关键词: AIDS prevention; AIDS/HIV problem; Accounting; Acquired Immunodeficiency Syndrome; Address; Affect; Autologous; Basic Science; CCR5 gene; CD4 Positive T Lymphocytes; Caring; Cell model; Cells; Cervix Mucus; Chronic; Clinical Treatment; Data; Educational workshop; Endometrial; Endometrium; Environment; Epithelial Cells; Epithelium; Female; Genome; Gonadal Steroid Hormones; HIV; HIV Receptors; HIV-1; Health; Heterosexuals; Human; Human Resources; Immunologics; Individual; Infection; Laboratories; Lamina Propria; Length; Luteal Phase; Mediating; Menstrual cycle; Microspheres; Modality; Modeling; Molecular Cloning; Molecular Virology; Mucosal Immunity; Mucous Membrane; Pathogenesis; Pathway interactions; Patient Care; Predisposition; Prevention; Prevention Research; Property; Recording of previous events; Reporter; Research; Research Priority; Research Project Grants; Research Proposals; Risk; Route; Sample Size; Seminal fluid; Sensitivity and Specificity; Services; Site; Source; Staging; T-Lymphocyte; Testing; Time; United States National Institutes of Health; Uterus; Vaccines; Veterans; Viral; Virion; Virus; Virus Diseases; Woman; Women' s Health; analytical method; fitness; health administration; innate immune function; innovation; male; microbicide; myometrium; pandemic disease; pathogen; programs; public health relevance; receptor; reproductive tract; research study; sperm cell; tool; transcytosis; transmission process; treatment trial; virus host interaction

 DESCRIPTION (provided by applicant): HIV/AIDS is a significant health risk to both male and female veterans, and is transmitted predominantly by heterosexual intercourse. A roadblock to the discovery of efficacious prevention modalities, including vaccines and microbicides, is the incomplete understanding of basic virus-host interactions that underlie the acquisition of mucosal HIV-1 infection in the female genital tract. Two recent NIH workshops on Mucosal Immunity and HIV Prevention highlighted under-explored fundamental questions that need to be addressed, including anatomical sites of early virus infection in the female reproductive tract (FRT), and the influence of endogenous sex hormones on the mucosal environment and susceptibility to HIV-1. We submit that the endometrial mucosa of the uterus constitutes a site of early HIV-1 infection. (i) The upper FRT (uFRT) is accessible to pathogens. The uterus is readily accessible to sperm, experimental microspheres, viral and bacterial pathogens, and possibly HIV-1 virions attached to sperm. (ii) The cervical mucus is an imperfect barrier and upward myometrium contractions likely promote delivery of semen content into the uFRT. (iii) The uFRT comprises an abundance of HIV-1 target cells beneath the epithelium lining. (iv) Cellular and innate immune functions in the uFRT are suppressed during the secretory phase of the menstrual cycle. (v) Previous studies on HIV-1 infection of UEC are. (vi) Innovative and transformative virologic strategies are available in our laboratory that will facilitate physiologically relevant studies using primary mucosal transmission models, including transmitted/founder (T/F) HIV-1 infectious molecular clones (IMC), and HIV- 1 "reporter genomes" that augment both sensitivity and specificity for dissecting early virus-host mucosal interactions. (vii) We present preliminary results that to our knowledge demonstrate for the first time UEC are susceptible to infection by T/F. Therefore, there exists essential virologic underpinnings and compelling rationale to further examine the uFRT as a potential site for the acquisition of mucosal HIV-1 infection and transmission. Our central hypothesis is that the uFRT mucosa is vulnerable to productive T/F HIV-1 infection. To test this hypothesis, we propose the following specific aims: (1) To determine the susceptibility of primary UEC to productive infection by T/F, chronic control and laboratory strains of HIV-1. Viruses expressing T/F envs from isogenic reporter proviral genomes will be studied, enabling sensitive and specific analyses of productive HIV-1 infection in a primary UEC model of virus infection. Importantly, chronic control viruses will be analyzed in parallel experiments to rigorously address the question of whether T/F viruses possess unique fitness properties for infecting potential target cells that exist in the uFRT mucosa. (2) To elucidate cis and trans infection pathways by which T/F HIV-1 spreads from primary UEC to endometrial CD4+ T lymphocytes. Mucosal CD4+ T cells isolated from endometrium will first be analyzed for their susceptibility to infection by T/F and control viruses to establish important underpinnings for subsequent experiments with UEC. The following experiments will elucidate susceptibilities of autologous CD4+ T cells to HIV-1 infection mediated by de novo produced virus from infected UEC or transcytosis of intact/infectious virus particles. The results will be analyzed to elucidate virus subtype- and strain-specific susceptibilities of UEC and CD4+ T lymphocytes to HIV-1 infection, and identify mechanisms/pathways of virus entry and correlations of these factors with menstrual cycle stage. Such findings promise to provide critical underpinnings for translational prevention research initiatives.

 描述（由申请人提供）： 艾滋病毒/艾滋病是男性和女性退伍军人的重大健康风险，并且主要通过异性恋性交传播。发现有效预防方式的障碍，包括疫苗和杀菌剂，是对基本病毒 - 宿主相互作用的不完全理解，这些相互作用是在女性生殖道中获得粘膜HIV-1感染的基础。最近两个关于粘膜免疫和艾滋病毒预防的NIH研讨会强调了需要解决的基本问题，包括女性生殖道早期病毒感染的解剖部位（FRT）和影响 内源性性恐怖在粘膜环境中以及对HIV-1的敏感性。我们认为子宫的子宫内膜粘膜构成了早期HIV-1感染的部位。 （i）病原体可以使用上部FRT（UFRT）。子宫很容易获得精子，实验性微球，病毒和细菌病原体以及附着在精子上的HIV-1病毒。 （ii）宫颈粘液是一种不完美的屏障，并且向上的子宫骨值收缩可能会促进精液含量递送到UFRT中。 （iii）UFRT在上皮衬里下方建立了大量的HIV-1靶细胞。 （iv）在月经周期的秘密阶段，UFRT中的细胞和先天免疫功能被抑制。 （v）先前关于UEC HIV-1感染的研究。 （vi）我们的实验室可以使用创新和变革性的病毒学策略，该策略将促进使用主要粘膜传播模型，包括传播/创建者（T/F）HIV-1感染性分子克隆（IMC）和HIV-1“记者基因组”，以增强敏感性和特异性早期的Virus Virus互动Mucs-sost-Mucs-sost-Mucs-sost Mucs-sost Mucs-sost-sost-sost-sost-sost sostistion niv-hiv-1感染性分子克隆（IMC）和HIV-1“记者基因组”。 （vii）我们提出了初步结果，据我们所知，UEC首次易受T/F感染的影响。因此，存在必要的病毒学基础和引人注目的理由，以进一步研究UFRT，作为获取粘膜HIV-1感染和传播的潜在部位。我们的中心假设是UFRT粘膜容易受到生产性T/F HIV-1感染的影响。为了检验这一假设，我们提出了以下特定目的：（1）确定原发性UEC对HIV-1的慢性控制和实验室菌株对生产性感染的敏感性。来自等生报道提供商基因组的T/F ENV的病毒将进行研究，从而在病毒感染的主要UEC模型中对生产性HIV-1感染进行了敏感和特定的分析。重要的是，将在平行实验中分析慢性控制病毒，以严格解决T/F病毒是否具有感染UFRT粘膜中存在潜在靶细胞的潜在适应性特性的问题。 （2）阐明T/F HIV-1从原发性UEC传播到子宫内膜CD4+ T淋巴细胞的顺式和反式感染途径。将首先分析从子宫内膜分离的粘膜CD4+ T细胞，以分析其通过T/F和对照病毒感染的敏感性，并为随后的UEC实验建立重要的基础。以下实验将阐明自体CD4+ T细胞对由从头开始产生的感染的UEC或完整/感染性病毒颗粒的转胞胞菌的病毒介导的HIV-1感染。将分析结果以阐明 UEC和CD4+ T淋巴细胞对HIV-1感染的病毒亚型和菌株特异性的敏感性，并确定这些因素的机制/途径以及这些因素与月经周期阶段的相关性。这样的发现有望为转化预防研究计划提供关键的基础。