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: 8892997
• 负责人: Siddappa N Byrareddy
• 金额: $ 86.69万
• 依托单位: EMORY UNIVERSITY
• 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 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8892997
• 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; Drug Formulations; Enhancers; Evolution; Fingerprint; Foundations; Future; Galactose Binding Lectin; Glycoproteins; HIV; HIV Envelope Protein gp120; HIV Infections; HIV vaccine; HIV-1; Health; Helper-Inducer T-Lymphocyte; Immune response; In Vitro; Infection; Interferon-alpha; Kinetics; Knowledge; Lectin; Ligands; Link; Macaca; Macaca mulatta; 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; Surface; T-Lymphocyte; Testing; Vaccine Design; Vaccines; Vagina; Variant; Vertebral column; Viral; Viral Antigens; Virus; base; cohort; deep sequencing; design; env Gene Products; glycosylation; immune function; in vivo; neutralizing antibody; nonhuman primate; particle; prevent; receptor; simian human immunodeficiency virus; sugar; tool; transmission process; vaginal 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（由适用提供）：对促进传播的病毒/宿主关系的精致理解以及感染后疾病进展速度的疾病进展速度的理解对于设计有效疫苗针对HIV-1至关重要。一个越来越多地被认为是至关重要和理解的参数是病毒env糖基化的作用，不仅在首选传播中，而且在病毒选择进入的受体以及其中和抗体的敏感性方面的能力。此外，数据现在积累的数据表明，多种细胞表面分子是糖基化的ENV蛋白的配体/受体，以及受糖基化和去糖基化病毒影响的细胞表面分子，可能有助于免疫反应的病毒发挥剂。这些问题只能使用适当的非人类灵长类动物模型来客观地解决，例如感染的猕猴感染了含有原发性HIV-1分离株的ENV，尤其是从传播/创始人病毒中研究ENV糖基化在首选传播中的作用。因此，我们计划使用从Zambian Cohort-Where-clade C病毒占主导地位的传输/创始人（T/F）病毒获得的一组HIV分子克隆探索Env Glycans的作用。我们已经生成了几种独特的工具/试剂，并组装了一支高度才华横溢的调查人员团队，并概述了一系列系统的逻辑设计的研究，该研究将重点关注来自T/F循环HIV循环HIV菌株中的Glycans的组成和布置，包括N-and O-Glycans，包括AIM 1。基于AIM级别的AIM级别。 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 tr​​ans-infection, sensitivity to IFN-α，抗α4ß7和针对精选杠杆样分子（Galectin-9，Siglecs，Mincle和CD200/200R）的mAb。然后，我们将选择病毒以在体内测试恒河猕猴的传播效率研究，主要基于可比的体外复制，但具有不同的聚糖含量（AIM 2）。在特定的目标3下，我们将定义是否是ENV糖基化的签名位点促进传播的促进，使用基于“贫穷”发射器的骨架，仅使用IAM 2B的基础病毒对应的依赖性SHIV，并使用重复的多重剂量多剂量的多种剂量多剂量内剂量内（IVAG）质疑模型。这些研究的结果将为未来的理性艾滋病毒疫苗设计奠定坚实的基础。