Discovery and Targeting of West Nile Virus Epitopes

西尼罗河病毒表位的发现和靶向

• 批准号: 8245072
• 负责人: William Hildebrand
• 金额: $ 115.97万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-09 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8245072
• 关键词: Academia; Animal Testing; Animals; Antibodies; Antiviral Agents; B-Lymphocytes; Biological Assay; C57BL/6 Mouse; Categories; Cell Lineage; Cell surface; Cells; Collaborations; Complex; Cytotoxic T-Lymphocytes; Defect; Development; Diagnosis; Diagnostic; Diamond; Epitopes; Family; Fever; Fiber; Flaviviridae; Future; Generations; Goals; HLA Antigens; Human; Human Development; Hybridomas; Immune response; Immune system; Immunotherapeutic agent; In Vitro; Individual; Industry; Infection; Intracellular Space; Laboratories; Lead; Life Cycle Stages; Logic; Lymphocyte; Mass Spectrum Analysis; Mediating; Meningoencephalitis; Mission; Monoclonal Antibodies; Mus; National Institute of Allergy and Infectious Disease; Neurologic; Pathogenesis; Peptide Fragments; Peptides; Physicians; Population; Principal Investigator; Production; Proteins; Research Personnel; Rodent; Sampling; Scanning; Seroprevalences; Specificity; Surface; Survivors; T-Cell Antigen Receptor Specificity; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Epitopes; Testing; Texas; Therapeutic; Therapeutic Intervention; Time; Transgenic Mice; Transgenic Organisms; Universities; Vaccines; Viral; Viral Proteins; Virulent; Virus; Virus Diseases; Washington; West Nile virus; base; catalyst; experience; fitness; in vivo; microbial; mouse model; neutralizing antibody; novel; novel strategies; pathogen; pre-clinical; programs; receptor; research clinical testing; response; therapeutic development; vaccine development; virus infection mechanism

DESCRIPTION (provided by applicant): Class I Human Leukocyte Antigens (HLA) is found on the surface of virtually every cell. Their function is to convey intracellular fitness to lymphocytes that cannot otherwise access intracellular spaces in order to detect viruses. Class I HLA molecules expose a viral infection by first sampling peptide fragments of proteins within the cell. Class I molecules then egress to the cell surface and display these intracellular protein snippets to T lymphocytes. Cytotoxic T lymphocytes which continuously scan class I HLA at the cell surface are able to single out class I HLA carrying virus-derived peptides and then destroy infected cells. Class I HLA reveals intracellular viruses by presenting viral peptide epitopes at the cell surface. Our laboratory is devoted to identifying those peptide epitopes unique to infected cells. We posit that the accurate discovery of viral epitopes unique to infected cells will facilitate the successful downstream development of viral therapeutics. To test this hypothesis, we propose to discover class I HLA presented peptide epitopes unique to West Nile Virus (WNV) infected cells in aim 1 of this project. We will identify WNV epitopes in multiple class I HLA using hollow fiber biorectors and mass spectroscopy. In order to initiate therapeutic development of the epitopes we find unique to WNV infected cells, in aim 2 we propose the generation of monoclonal antibodies to WNV/HLA class I complexes that distinguish infected cells. The development of mAb to WNV/HLA complexes in aim 2 will proceed through collaboration with Receptor Logic, a biotech firm proficient in the production of mAb to HLA/peptide complexes. The overall objective of this project is to therapeutically target WNV infected cells with mAb, and in aim 3 we will test these mAb for their ability to destroy WNV infected cells in vivo. HLA transgenic mice will be infected with WNV, mAb to WNV peptide/HLA complexes will be passively administered, and the antibody-mediated destruction of virus-infected cells will be evaluated.

描述（由申请人提供）： I 类人类白细胞抗原 (HLA) 几乎存在于每个细胞的表面。它们的功能是将细胞内的适应性传递给淋巴细胞，否则这些淋巴细胞无法进入细胞内空间以检测病毒。 I 类 HLA 分子首先通过对细胞内蛋白质的肽片段进行采样来暴露病毒感染。然后 I 类分子逸出到细胞表面，并将这些细胞内蛋白质片段展示给 T 淋巴细胞。细胞毒性T淋巴细胞在细胞表面连续扫描I类HLA，能够挑出携带病毒衍生肽的I类HLA，然后破坏受感染的细胞。 I 类 HLA 通过在细胞表面呈现病毒肽表位来揭示细胞内病毒。我们的实验室致力于鉴定受感染细胞特有的肽表位。我们认为，准确发现受感染细胞特有的病毒表位将有助于病毒疗法下游的成功开发。为了检验这一假设，我们建议在本项目的目标 1 中发现西尼罗河病毒 (WNV) 感染细胞特有的 I 类 HLA 呈递肽表位。我们将使用中空纤维生物反应器和质谱来识别多个 I 类 HLA 中的 WNV 表位。为了启动我们发现的 WNV 感染细胞特有的表位的治疗开发，在目标 2 中，我们建议生成针对 WNV/HLA I 类复合物的单克隆抗体，以区分受感染的细胞。目标 2 中针对 WNV/HLA 复合物的 mAb 开发将通过与 Receptor Logic 合作进行，Receptor Logic 是一家精通生产针对 HLA/肽复合物的 mAb 的生物技术公司。该项目的总体目标是用 mAb 治疗性靶向 WNV 感染细胞，在目标 3 中，我们将测试这些 mAb 在体内破坏 WNV 感染细胞的能力。 HLA转基因小鼠将感染WNV，被动施用针对WNV肽/HLA复合物的mAb，并评估抗体介导的病毒感染细胞的破坏。