Role of HIV Env glycosylation in mucosal transmission

HIV Env 糖基化在粘膜传播中的作用

• 批准号: 9294954
• 负责人: Siddappa N Byrareddy
• 金额: $ 70.55万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-16 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9294954
• 关键词: Acute; Address; Affinity; Antibodies; Antigen-Presenting Cells; Antigens; Area; Binding; CD 200; Cell Line; Cell surface; Cells; Chronic; Competence; Data; Dendritic Cells; Disease Progression; Dose; Enhancers; Evolution; Fingerprint; Formulation; Foundations; Future; Galactose Binding Lectin; Glycoproteins; HIV; HIV Envelope Protein gp120; HIV Infections; HIV vaccine; HIV-1; Helper-Inducer T-Lymphocyte; Immune response; In Vitro; Infection; Interferon-alpha; Kinetics; Knowledge; Lectin; Ligands; Link; Macaca; Macaca mulatta; Membrane Proteins; Modeling; Molecular Cloning; Mucous Membrane; Mutate; Pathogenesis; Play; Polysaccharides; Positioning Attribute; Predisposition; Property; Proteins; Reagent; Receptor Cell; Recombinants; Research Design; Research Personnel; Resources; Role; SIV; Series; Site; Source; T-Lymphocyte; Talents; Testing; Vaccine Design; Vaccines; Variant; Vertebral column; Viral; Viral Antigens; Viral Pathogenesis; Virus; Virus Replication; base; cohort; deep sequencing; design; env Gene Products; glycosylation; immune function; in vivo; neutralizing antibody; nonhuman primate; particle; prevent; public health relevance; receptor; sialic acid binding Ig-like lectin; simian human immunodeficiency virus; sugar; tool; transmission process; vaginal transmission; viral transmission

DESCRIPTION (provided by applicant): A refined understanding of virus/host relationships that promote transmission and those that contribute to the rate of disease progression following infection is critical for the design of effective vaccines against HIV-1. One parameter that is increasingly recognized as being critical and understudied is the role of viral env glycosylation not only in preferential transmission but also in the ability of the virus to select receptors for entry and for its susceptibility to neutralizing antibodies. In addition, data is now accumulating o show that a variety of cell surface molecules that serve as ligands/receptors for the glycosylated env proteins and those that are influenced by glycosylated versus de-glycosylated viruses, could contribute to the quality of immune response the virus engenders. These issues can only be objectively addressed using appropriate nonhuman primate models such as macaques infected with SHIVs that contain the env from primary HIV-1 isolates, particularly from transmitted/founder viruses to study the roles of env glycosylation in preferential transmission. Therefore, we plan to explore the role of env glycans using sets of HIV molecular clones obtained from transmitted/founder (T/F) viruses from a Zambian cohort-where clade C viruses predominate. We have generated several unique tools/reagents and assembled a highly talented team of Investigators and a series of systematic logically designed studies are outlined which will first focus on the composition and arrangement of glycans in paired clones from T/F circulating HIV strains including N-and O-glycans as outlined in aim 1. Based on glycan profiles, we proposed to select a) a highly glycosylated, b) low level of env glycosylated viruses from transmitter and c) as control, a founder virus to prepare corresponding replication competent Simian Human Immunodeficiency viruses (SHIV) for further advanced studies, which include detailed in vitro characterization of replication kinetics in primary cells and cell lines, glycan content, particle envelope glycoprotein content, dendritic cell (DC) capture, DC-T cell trans-infection, sensitivity to IFN-α, anti-α4ß7 and blocking mAbs against select lectin-like molecules (Galectin-9, SIGLECs, MINCLE, and CD200/200R). Then we will select the viruses to test in vivo for transmission efficiency studies in rhesus macaques based primarily on comparable in vitro replication but differing glycan content (aim 2). Under specific aim 3, we will define whethe it is the signature site(s) of Env glycosylation that facilitates transmission, using isogenic SHIV based on the backbone of the "poor" transmitter and mutating only those select residues corresponding to the founder virus from aim 2b, using a repeated multiple-low-dose Intra-Vaginal (IVAG) challenge model. Results of these studies will provide a strong foundation for future rational HIV vaccine design.

描述（由申请人提供）：对促进传播的病毒/宿主关系以及对感染后疾病进展速度的贡献的病毒/宿主关系的精确理解对于设计有效的HIV-1疫苗至关重要。越来越多地被认为是关键的和未充分研究的一个参数是病毒env糖基化的作用，不仅在优先传输，而且在病毒选择受体的能力进入和其对中和抗体的敏感性。此外，现在积累的数据表明，作为糖基化env蛋白的配体/受体的各种细胞表面分子以及受糖基化与去糖基化病毒影响的那些细胞表面分子可能有助于病毒产生的免疫应答的质量。这些问题只能使用适当的非人灵长类动物模型来客观地解决，例如感染含有来自主要HIV-1分离株（特别是来自传播/创始者病毒）的env的SHIV的猕猴，以研究env糖基化在优先传播中的作用。因此，我们计划探索env聚糖的作用，从传播/创始人（T/F）病毒从赞比亚cohort，其中C型病毒占主导地位的艾滋病病毒分子克隆集。我们已经生成了几种独特的工具/试剂，并组建了一支非常有才华的研究团队，概述了一系列系统的逻辑设计研究，这些研究将首先关注T/F循环HIV毒株配对克隆中聚糖的组成和排列，包括目标1中概述的N-和O-聚糖。基于聚糖谱，我们建议选择a）高度糖基化，B）低水平env糖基化的病毒和c）作为对照，建立病毒以制备相应的复制能力的猴人免疫缺陷病毒（SHIV）用于进一步的进一步研究，包括详细的体外表征在原代细胞和细胞系中的复制动力学，聚糖含量，颗粒包膜糖蛋白含量，树突状细胞（DC）捕获、DC-T细胞转感染、对IFN-α的敏感性、抗α 4 β 7和针对选择的凝集素样分子（半乳糖凝集素-9、SIGLEC、MINCLE和CD 200/200 R）的阻断mAb。然后，我们将选择病毒进行体内检测，主要基于体外复制的可比性，但聚糖含量不同（目的2），用于恒河猴中的传播效率研究。在具体目标3下，我们将使用基于“弱”传递物骨架的同基因SHIV并使用重复的多次低剂量阴道内（IVAG）攻击模型仅突变对应于目标2b的创始病毒的那些选择残基，来确定是否是Env糖基化的特征位点促进传播。这些研究结果将为今后合理设计HIV疫苗提供坚实的基础。