Mechanisms of Papillomavirus Neutralization

乳头瘤病毒中和机制

基本信息

批准号：
7471412
负责人：
Richard Bruce Roden
金额：
$ 28.04万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-07-19 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7471412
关键词：
Address Animals Antibodies Binding Binding Sites Biochemical Genetics Capsid Capsid Proteins Cell physiology Cell surface Cells Conflict (Psychology)Cryoelectron Microscopy Data Developed Countries Developing Countries Development Dissection Epitopes Escherichia coli Experimental Models Gardasil Genotype Goals Gold Human Human Papillomavirus Human papilloma virus infection Human papillomavirus 16 Immunity In Vitro Infection L1 viral capsid protein Life Cycle Stages Literature Location Malignant neoplasm of cervix uteri Mediating Minor Modeling Molecular Virology Monoclonal Antibodies Oncogenic Oryctolagus cuniculus Pan Genus Papillomavirus Papillomavirus Infections Phenotype Prevention Preventive Process Proteins Range Reporting Research Personnel Screening procedure Serotyping Standards of Weights and Measures Vaccination Vaccines Viral Viral Genome Virion Virus Virus Diseases Virus-like particle base image reconstruction monolayer mutant neutralizing antibody neutralizing monoclonal antibodies particle prevent programs syntaxin 18 three dimensional structure trafficking transmission process

项目摘要

DESCRIPTION (provided by applicant): Persistent infection by one of >15 oncogenic Human papillomavirus (HPV) types is a necessary cause of cervical cancer. Papillomavirus has only two capsid proteins; L1 and L2. Although L1 is known to form the capsid and mediates initial binding to cells, the functions of L2 remain to be determined. Whereas vaccination with L1 virus-like particles induces type-restricted immunity, we show that vaccination of animals with L2 provides broad immunity from viral challenge via neutralizing antibodies. Therefore, our long-term goal is the rational development of a single, inexpensive pan-oncogenic HPV preventive vaccine through dissection of mechanisms relating to L2 function in the infectious process and L2-dependent neutralization. We find that L2 is required for appropriate intracellular trafficking and binds to the cellular proteins syntaxin-18 and VAV-2. L2 neutralizing antibodies permit virus binding to cell monolayers, but their epitopes overlap the binding sites of interacting cellular proteins, suggesting: L2 antibody-dependent neutralization occurs by preventing interaction between L2 and key cellular proteins that mediate papillomavirus trafficking. To address this hypothesis we propose Specific aim 1: Characterize the effect of L2-specific neutralizing monoclonal antibodies upon the trafficking of virions and interaction with cellular proteins during infection, and Specific aim 2: Generate fine mutants within HPV L2 that prevent interaction with cellular proteins and determine their influence upon the particle to infectivity ratio of HPV pseudotype virions. Conflicting reports of L2 function in the literature likely reflect differences in the plethora of experimental models applied to study the molecular virology of papillomavirus. The organotypic raft model is the 'gold standard' that most closely replicates the entire HPV life cycle. Therefore we propose Specific aim 3: Validate the neutralizing activity of the L2 monoclonal antibodies and the phenotype of mutant HPV16 L2 in the context of the viral genome using the organotypic raft model of the complete viral life cycle. Finally we propose Specific Aim 4: Determine the three dimensional structure of HPV16 bound to neutralizing and non-neutralizing L2-specific monoclonal antibodies by cryo- electron microscopy and image reconstruction. These studies will identify conserved interactions between L2 and cellular proteins that are critical to infection and targeted by protective antibodies.

描述（由申请人提供）：持续感染> 15个致癌人乳头瘤病毒（HPV）类型是宫颈癌的必要原因。乳头瘤病毒只有两个衣壳蛋白。 L1和L2。尽管已知L1形成衣壳并介导初始结合与细胞，但L2的功能仍有待确定。尽管L1病毒样颗粒接种疫苗会诱导类型限制的免疫力，但我们表明，具有L2的动物的疫苗接种通过中和抗体可通过中和抗体提供广泛的免疫力。因此，我们的长期目标是通过解剖与传染过程中与L2功能相关的机制和L2依赖性中和的机制，从而合理地发展了一个廉价的，廉价的泛氧化HPV预防性疫苗。我们发现L2是适当的细胞内运输所必需的，并与细胞蛋白语法-18和VAV-2结合。 L2中和抗体允许病毒与细胞单层结合，但是它们的表位与相互作用的细胞蛋白的结合位点重叠，这表明：L2抗体依赖性中和通过预防L2与介导乳头虫贩运的钥匙细胞蛋白之间的相互作用发生。 To address this hypothesis we propose Specific aim 1: Characterize the effect of L2-specific neutralizing monoclonal antibodies upon the trafficking of virions and interaction with cellular proteins during infection, and Specific aim 2: Generate fine mutants within HPV L2 that prevent interaction with cellular proteins and determine their influence upon the particle to infectivity ratio of HPV pseudotype virions.文献中L2功能的报道冲突可能反映了用于研究乳头瘤病毒分子病毒学的许多实验模型的差异。器官型筏模型是“黄金标准”，最紧密地复制整个HPV生命周期。因此，我们提出了特定的目标3：使用完整病毒生命周期的器官型RAFT模型，在病毒基因组的背景下，在病毒基因组的背景下验证L2单克隆抗体的中和活性和突变体HPV16 L2的表型。最后，我们提出了特定的目标4：确定与冷冻显微镜和图像重建结合的HPV16的三维结构。这些研究将确定L2与细胞蛋白之间的保守相互作用，这些相互作用对感染至关重要，并且由保护性抗体靶向。