Determinants of HIV transmission fitness

假设：HIV包膜糖蛋白(Env)上有一个具有特定糖组成的最小糖膜-- 1可能是确定新感染所必需的。在艾滋病毒中选择了一种新的感染方式，即多糖 根据传播的方式和容器(例如直肠，阴茎， 包皮、阴道和宫颈组织在性传播中)以及细胞因子/趋化因子环境和 微生物组。了解在这些微环境中传播的HIV-1上的多糖组成将 改进基于体液的HIV-1疫苗的设计和效果。 在过去的15年里，这项R01将艾滋病毒疾病参数与T细胞中复制艾滋病毒的适合性进行了比较， 巨噬细胞，树突状细胞共培养，以及各种体外非粘膜组织。我们已经证明了 患者来源的HIV的竞争性复制适合性是致病机制的替代，并且在缺乏 治疗与疾病进展有很强的相关性。然而，我们发表的工作表明， HIV-1初级分离株的“致病性”和“传播性”适合度相当不同。使用不同的条件 人外植体阴茎和宫颈(ENC)组织接触20多种HIV-1来源的不同混合物 从急性和慢性疾病来看，我们现在对影响艾滋病毒传播的因素有了更清晰的了解。我们 观察到慢性HIV-1在粘膜(阴茎和宫颈)组织中的“陷阱”复制，而急性HIV-1 分离株通过组织(可能通过树突状细胞)感染Th17或Th1 CD4+T细胞。HIV-1，即 “逃逸”的粘膜组织通常对C型凝集素的亲和力较低，N连接的糖基化位点较少。 为了加强这些研究，我们与程序员和生物信息学专家密切合作，开发出可靠的 NGS分析管道，以确定多病毒竞争的传播适宜性，最近，我们 优化了分析gp120多聚糖的EIS-MS技术。 我们提出了四个高度相关的目标来检验我们的假设： 具体目的1.宫颈腔内(ENC)和血浆样本中HIV-1的体内分析和决定因素 新的艾滋病毒传播。检测新感染妇女鼻咽癌和血液中的env基因序列特征 ENC中感染艾滋病毒-1但尚未血清转换的妇女也是如此。 具体目标2.表征血液中传播/创立者(TF)HIV变异株与 HIV-1在女性和男性生殖道中的传播：体外研究。措施被困 人体阴茎、包皮、宫颈和阴道移植模型中组织内复制与传播的比较。 具体目的3.测定凝集素结合亲和力并分析gp120上的糖基化模式 用新的gp120纯化技术和EIS-MS检测高传播性转铁蛋白HIV-1的环境。 特定目的4.修饰TF和ENC病毒的葡聚糖并确定最适的HIV环境 使用目标2和目标3中描述的相同方法实现艾滋病毒从男性到女性的传播。

Hypothesis: A minimal glycan shield with specific sugar composition on the envelope glycoprotein (Env) of HIV- 1 may be necessary for establishing new infections. In the HIV “selected” for a new infection, glycan content/composition on Env could differ based on the mode and receptacle of transmission (e.g. rectal, penile, foreskin, vaginal, and cervical tissue in sexual transmission) as well as the cytokine/chemokine milieu and microbiome. Understanding glycan composition on the transmitted HIV-1 in these microenvironments would improve the design and effectiveness of humoral-based HIV-1 vaccine. For the past 15 years, this R01 has compared HIV disease parameters to replicative HIV fitness in T cells, macrophages, dendritic cell cocultures, and various ex vivo, non-mucosal tissues. We have shown that competitive replicative fitness of patient-derived HIV is a surrogate of pathogenesis and in the absence of treatment, was a strong correlate of disease progression. However, our published work indicates that “pathogenic” and “transmission” fitness of primary HIV-1 isolates are quite distinct. Using varying conditions with human explant penile and endocervical (EnC) tissues exposed to over 20 different mixtures of HIV-1 derived from acute and chronic disease, we now have a clearer picture of factors impacting HIV transmission. We observed “trapped” replication of chronic HIV-1 in mucosal (penile and cervical) tissue while the acute HIV-1 isolates were carried through the tissue (possibly via DCs) to infect Th17 or Th1 CD4+ T cells. HIV-1 that “escapes” mucosal tissues typically have lower affinity for C type lectins and fewer N linked glycosylation sites. To enhance these studies we worked closely with programmers and bioinformatic specialists to develop robust NGS analysis pipelines to determine transmission fitness from multivirus competitions and more recently, we optimized EIS-MS techniques to analyze gp120 glycans. We have proposed four highly inter-related aims to test our hypotheses: Specific Aim 1. In vivo analyses of HIV-1 in endocervical (EnC) and plasma samples and determinants of new HIV transmissions. Examine Env sequence features in the EnC and blood of newly infected women as well as in women with HIV-1 infection in the EnC but who have not seroconverted. Specific Aim 2. Characterizing the properties of transmitted/founder (TF) HIV variant in blood versus the transmitted HIV-1 population in the female and male genital tract: Ex vivo study. Measure trapped replication in the tissue versus transmission in human penile, foreskin, cervical, and vaginal explant model. Specific Aim 3. To measure lectin binding affinity and to analyze the glycosylation pattern on the gp120 Env of highly transmissible TF HIV-1 using new gp120 purification techniques and EIS-MS. Specific Aim 4. To modify the glycans on TF and EnC viruses and to determine optimal HIV Env for male to female HIV transmissions using the same methods described in aims 2 and 3.