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/艾滋病对男女退伍军人都是一个重大的健康风险，主要通过异性性交传播。发现有效的预防方式，包括疫苗和杀微生物剂的一个障碍是对女性生殖道粘膜HIV-1感染的基础病毒-宿主相互作用的不完全理解。最近两次NIH关于粘液免疫和艾滋病预防的研讨会强调了需要解决的未充分探索的基本问题，包括女性生殖道（FRT）早期病毒感染的解剖部位，以及病毒对女性生殖道（FRT）的影响。 内源性性激素对粘膜环境和HIV-1易感性的影响。我们认为子宫内膜粘膜是HIV-1早期感染的一个部位。(i)上FRT（uFRT）可接近病原体。精子、实验性微球、病毒和细菌病原体以及可能附着在精子上的HIV-1病毒体都容易进入子宫。(ii)宫颈粘液是一个不完善的屏障，子宫肌层向上收缩可能促进精液内容物进入uFRT。(iii)uFRT在上皮衬里下包含丰富的HIV-1靶细胞。(iv)uFRT中的细胞和先天免疫功能在月经周期的分泌期受到抑制。(v)HIV-1感染UEC的研究进展如下。(vi)创新和变革的病毒学策略，可在我们的实验室，将促进生理学相关的研究，使用主要的粘膜传播模型，包括传输/创始人（T/F）HIV-1感染性分子克隆（IMC），和HIV- 1的“报告基因组”，增加了解剖早期病毒-宿主粘膜相互作用的灵敏度和特异性。(vii)我们目前的初步结果表明，据我们所知，第一次UEC是易受感染的T/F。因此，存在必要的病毒学基础和令人信服的理由，以进一步研究uFRT作为获得粘膜HIV-1感染和传播的潜在位点。我们的中心假设是uFRT粘膜易受生产性T/F HIV-1感染。为了验证这一假设，我们提出了以下具体目标：（1）确定原发性UEC对T/F、慢性对照和实验室HIV-1株的生产性感染的敏感性。将研究从同基因报告基因前病毒基因组表达T/F envs的病毒，从而能够在病毒感染的主要UEC模型中对生产性HIV-1感染进行敏感和特异性分析。重要的是，将在平行实验中分析慢性对照病毒，以严格解决T/F病毒是否具有感染uFRT粘膜中存在的潜在靶细胞的独特适应性的问题。(2)阐明T/F HIV-1从原发性子宫内膜异位症子宫内膜上皮细胞（UEC）向子宫内膜CD 4 + T淋巴细胞传播的顺式和反式感染途径。将首先分析从子宫内膜分离的粘液CD 4 + T细胞对T/F和对照病毒感染的敏感性，以建立后续UEC实验的重要基础。以下实验将阐明自体CD 4 + T细胞对HIV-1感染的易感性，所述HIV-1感染由来自感染的UEC的从头产生的病毒或完整/感染性病毒颗粒的转胞吞作用介导。将对结果进行分析， UEC和CD 4 + T淋巴细胞对HIV-1感染的病毒亚型和毒株特异性易感性，并确定病毒进入的机制/途径以及这些因素与月经周期阶段的相关性。这些发现有望为转化预防研究举措提供关键基础。