Dichotomy of HIV-Sugar with Vaginal Microbes

项目摘要 这个提议是建立在PI Mak实验室的一个新发现之上的， 特异性、可操纵的糖-糖是HIV的附着因子。这种相互作用发生在艾滋病毒 在受体接合之前，寡聚甘露糖（oligoman）和细胞N-乙酰葡糖胺（GlcNAc）。的 跨生命和非生命（病毒）领域的聚糖生物学的保守性导致了我们目前的提议 研究这种寡聚体-GlcNAc（或基于寡聚体的）相互作用是否可能支持一些动态- 相互作用：（i）阴道微生物;（ii）HIV;和（iii）它们的人类宿主。我们的发现是艾滋病病毒加倍， 使用其富含寡聚体的HIV包膜（Env）聚糖屏蔽作为附着因子可以提供一种通用原理 至少部分地解释：（i）阴道分泌物如何保护免受艾滋病毒传播;以及（ii）如何 阴道淋病奈瑟菌增加了艾滋病毒传播的风险。很多阴道微生物都有甘露糖- 结合蛋白和GlcNAc的细菌表面，我们的假设是，富集寡聚N- HIV Env上的聚糖作为分子Velcro与甘露糖结合蛋白和/或GlcNAc糖相互作用 在生殖器微生物的表面。我们进一步假设，这些生殖器微生物的生物学特性 要么帮助诱捕艾滋病毒，使其无法感染宿主，要么充当信使服务，将艾滋病毒运送到宿主体内。 主持人我们将结合使用超微结构成像技术，生物物理方法，分子病毒学 技术、飞行时间流式细胞术（CyTOF）-成像、聚糖基质辅助激光解吸电离- 质谱成像（MALDI-MSI）和基于组织培养的上皮细胞模型系统，以检查 通过基于寡聚体的结合通过两个特定目的传播HIV。在目标1中，我们将定义阴道是否 HIV通过寡聚体捕获来限制HIV传播。在目标2中，我们将评估艾滋病毒是否会在 通过寡聚体介导的与淋病奈瑟菌的结合而形成上皮下屏障。成功完成本 该提案可能会导致概念数据的证明，即基于简单寡聚体的机制可以解释 淋病奈瑟菌如何保护和如何增强HIV传播的相反能力。 我们还将使用来自ATCC的预设阴道微生物参考群体来评估这些基于聚糖的 在人口层面的互动。我们将对阴道微生物组进行随访分析， 已建立并具有临床相关性的南非艾滋病毒队列。数据显示，基于寡聚体的相互作用是一种 艾滋病毒性传播的一个促成因素可能为开发一种实用的口服 使用优化形式的GlcNAc或甘露糖递送自我施用的HIV缓解策略，鉴于这些 母体单糖是广泛可获得的、无毒的、非处方的营养品。

PROJECT SUMMARY This proposal is built upon a novel discovery from the PI Mak laboratory showing that a ‘non-electrostatic, specific, manipulatable, sugar-sugar’ is an attachment factor for HIV. This interaction occurs between HIV oligomannose (oligoman) and cellular N-acetylglucosamine (GlcNAc) prior to receptor engagement. The conservation of glycan biology across domains of life and the non-living (viruses) has led to our current proposal to investigate whether this oligoman-GlcNAc (or oligoman-based) interaction may underpin some of the dynamic- interplays amongst: (i) vaginal microbes; (ii) HIV; and (iii) their human host. Our findings that HIV double up to use its oligoman enriched HIV envelope (Env) glycan shield as an attachment factor may offer a general principle to account, at least in part, for: (i) how vaginal lactobacillus offers protection from HIV transmission; and (ii) how vaginal Neisseria gonorrhoeae increases the risk of HIV transmission. As many vaginal microbes have mannose- binding proteins and GlcNAc on their bacterial surface, our hypothesis is that the enrichment of oligoman N- glycans on HIV Env acts as a molecular Velcro to interact with mannose-binding proteins and/or GlcNAc sugars on the surface of genital microbes. We further hypothesize that the biology of these genital microbes would either helps trap HIV to deny their access to infect the host or acts as a courier service to deliver HIV into the host. We will use a combination of ultrastructural imaging techniques, biophysical approaches, molecular virology techniques, cytometry by time of flight (CyTOF) -imaging, glycan matrix-assisted laser desorption ionization - mass spectrometry imaging (MALDI-MSI), and tissue culture-based epithelial cell model system to examine the transmission of HIV via oligoman-based binding via two specific aims. In Aim1, we will define whether vaginal lactobacillus limits HIV transmission via oligoman-based trapping. In Aim2, we will assess if HIV hitch-hike across sub-epithelial barrier via oligoman-mediated bindings with N.gonorrhoeae. Successful completion of this proposal may result in proof of concept data that a simple oligoman-based mechanism could account for the opposing abilities of how lactobacillus protect and how N.gonorrhoeae enhance HIV transmission, respectively. We will also use pre-set vaginal microbes reference population from ATCC to evaluate these glycan-based interaction at the population level. We will perform follow-up analyses with vaginal microbiomes derived from established and clinically relevant South African HIV cohort. Data showing oligoman-based interaction is a contributing factor to sexual transmission of HIV may offer new hope for the development of a practical, orally delivery self-administrated HIV mitigation strategy using optimised versions of GlcNAc or mannose, given these parental monosaccharides are widely available, non-toxic, off-the counter, nutraceuticals.