Synthetic Variable Domain Glycopeptides for Neutralizing Epitope Characterization

用于中和表位表征的合成可变结构域糖肽

• 批准号: 8418151
• 负责人: LAI-XI WANG
• 金额: $ 23.03万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-05 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8418151
• 关键词: AIDS/HIV problem; Affinity; Amino Acids; Antibodies; Antigens; Binding; Biotin; Complex; Constitution; Crystallization; Data; Enzyme-Linked Immunosorbent Assay; Epidemic; Epitope Mapping; Epitopes; Future; Glycopeptides; Glycoproteins; Goals; HIV; HIV Envelope Protein gp120; HIV-1; Health; Heterogeneity; Human; Immobilization; Kinetics; Measures; Methods; Molecular Conformation; N-Glycosylation Site; Nature; Oligosaccharides; Peptides; Polysaccharides; Reporting; Research; Roentgen Rays; Screening procedure; Series; Serum; Site; Structure; Surface Plasmon Resonance; Technology; Testing; Vaccine Design; Vaccines; base; design; glycosylation; insight; mannosyl(5)-N-acetyl(2)-glucose; mannosyl(9)-N-acetylglucosamine2; neutralizing antibody; novel; research study; social

DESCRIPTION (provided by applicant): Identification and characterization of broadly neutralizing epitopes on the HIV-1 envelope are important steps in the design of an effective HIV-1 vaccine. Recently, a new class of broadly neutralizing antibodies (bNAbs), including PG9, PG16, and the PGT class antibodies, has been isolated from HIV-infected "elite controllers". These bNAbs neutralize primary HIV-1 strains with remarkable breadth and potency. A common feature of antigen recognition by these bNAbs is that they all target glycan-dependent quaternary epitopes at the V1/V2 and/or V3 regions of gp120. Recent X-ray crystal structural studies indicate that PG9 binds to N-glycans at N160 and N156 in the context of V1/V2 domain, and PGT128 recognizes conserved N-glycans at N322 and N301 sites in the context of V3 domain. However, the precise nature of the neutralizing epitopes, particularly the fine structures of the N-glycans at N156 and N301 remains to be characterized. Further mapping of the epitopes is complicated by the complexity and heterogeneity of glycosylation of HIV-1 gp120. We hypothesize that unique V1/V2 and V3 glycopeptides constitute the neutralizing epitopes for these bNAbs. To test this hypothesis, we will perform experiments described in two specific aims. Aim 1 is to design and synthesize cyclic V1/V2 and V3 HIV-1 glycopeptides with defined N-glycans being attached at the conserved N- glycosylation sites, by a novel chemoenzymatic method. Aim 2 is to characterize antigen recognition by the neutralizing antibodies through binding and structural studies with the synthetic glycopeptides. In addition, the synthetic glycopeptides will be used to detect glycan- dependent, V1/V2 and V3-specific neutralizing antibodies in sera from HIV-infected "non- progressors". These studies are likely to provide important insights for HIV-1 vaccine design. PUBLIC HEALTH RELEVANCE: HIV/AIDS is a serious global epidemic that threatens human health and social stability. The proposed research aims to characterize the neutralizing epitopes of broadly neutralizing antibodies, which will provide important insights in HIV-1 vaccine design.

描述（由申请人提供）：HIV-1包膜上广泛中和表位的识别和表征是设计有效HIV-1疫苗的重要步骤。最近，已经从HIV感染的“精英控制器”中分离出了一类新的广泛中和抗体（BNAB），包括PG9，PG16和PGT类抗体。这些bnabs以显着的宽度和效力中和原发性HIV-1菌株。这些BNAB的抗原识别的一个共同特征是，它们都靶向GP120的V1/V2和/或V3区域，靶向依赖聚糖的季度表位。最近的X射线晶体结构研究表明，在V1/V2结构域的背景下，PG9与N160和N156的N-聚糖结合，PGT128在V3域的上下文中识别N322和N301位点的保守N-聚糖。然而，中和表位的确切性质，尤其是N156和N301时N-聚糖的精细结构。 HIV-1 GP120的糖基化的复杂性和异质性使表位的进一步映射变得复杂。我们假设独特的V1/V2和V3糖肽构成了这些BNAB的中和表位。为了检验这一假设，我们将执行两个特定目标中描述的实验。 AIM 1是通过一种新型的化学酶方法设计和合成具有定义的N-聚糖的环状V1/V2和V3 HIV-1糖肽。目的2是通过与合成糖肽结合和结构研究中和结构研究来表征抗原识别的。此外，合成糖肽将用于检测来自HIV感染的“非奔跑者”的血清中血清中的V1/V2和V3特异性中和抗体。这些研究可能会为HIV-1疫苗设计提供重要的见解。 公共卫生相关性：艾滋病毒/艾滋病是一种严重的全球流行病，威胁到人类健康和社会稳定。拟议的研究旨在表征广泛中和抗体的中和表位，这将为HIV-1疫苗设计提供重要的见解。