Mechanisms of Papillomavirus Neutralization

乳头瘤病毒中和机制

基本信息

批准号：
8694133
负责人：
Richard Bruce Roden
金额：
$ 29.16万
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 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8694133
关键词：
Accounting Adenoviruses Affinity Alzheimer&apos s Disease Amyloid beta-Protein Precursor Animal Model Anogenital cancer Anogenital venereal warts Antibodies Antibody Degradation Antigens Antiviral Agents Binding Cancer Etiology Cell Nucleus Cells Cervical Cessation of life Cleaved cell Collaborations Cytoplasm Developing Countries Disease Epidermodysplasia Verruciformis Epitopes Escherichia coli Exhibits Fc Receptor Funding Genome Goals HIV HPV-High Risk Head and Neck Cancer Health Health Care Costs Hour Human Papilloma Virus Vaccine Human Papillomavirus Human papilloma virus infection Immune Immunoglobulin G In Vitro India Infection L2 viral capsid protein Licensing Link Low Income Population Low risk HPV Malignant Neoplasms Malignant neoplasm of cervix uteri Mediating Membrane Membrane Proteins Minor Morbidity - disease rate Normal Cell Oncogenic Papillomavirus Patients Play Price Proteolysis Role Senile Plaques Side Signal Transduction Site Skin Skin Carcinoma Sum Testing Transmembrane Domain Ubiquitination Vaccination Vaccine Antigen Vaccines Viral Viral Genome Virus Virus-like particle Woman base cost extracellular gamma secretase inhibitor/antagonist late endosome neurotoxic neutralizing antibody prevent prophylactic secretase trafficking uptake

项目摘要

DESCRIPTION (provided by applicant): Our overall goal is to understand how L2-specific antibodies effect broad protection against HPV infection. Here, we dissect why L2 and intra-membranous cleavage by gamma-secretase are required for infection, and how antibodies to L2 potently protect by neutralization at multiple steps during infection. In the initial funding perio we made the surprising observation that gamma-secretase is absolutely required for infection by HPV (Karanam et al., 2010), but only once HPV has begun to disassemble in the late endosome and gamma-secretase's proteolytic activity is required for escape from late endosomes of L2 in association with the viral genome. Recent findings suggest that L2 has a transmembrane-like domain between residues 45-67 that is essential for infection. We propose that the putative transmembrane region pierces the endosomal membrane, and gamma-secretase cleavage releases L2's carboxy terminus to bring the viral genome to the nucleus. Two key neutralizing epitopes in L2 reside either side of L2's putative transmembrane domain. Addition of L2 antibodies even several hours after the binding of virus to cells is neutralizing, suggesting blockade occurs late in infection. Therefore, we propose: Hypothesis 1: cleavage of L2 within its putative transmembrane region by gamma-secretase is critical for endosomal escape and is inhibited by L2-specific neutralizing antibodies. Specific Aim 1A: To determine the residues of L2 required for membrane insertion and whether they contribute to infectivity by facilitating endosomal escape. Specific Aim 1B: To determine the functional role and site of gamma secretase cleavage during HPV infection and the impact of L2-specific neutralizing antibodies and gamma secretase inhibitors on endosomal escape. Antibodies have generally been assumed to effect protection in a purely extracellular manner, as they are not generally found in the cytoplasm. Recently however, a TRIM21-dependent mechanism of antibody- dependent intracellular neutralization (ADIN) has been described. TRIM21 is a cytoplasmic high affinity Fc- receptor for the Fc of virally-associated antibody in the cytoplasm, triggering degradation of the antibody-bound virus. Our new findings suggest that ADIN may be important last line of defense in protection mediated by L2 antibodies. Hypothesis 2: the Fc-region of L2-specific neutralizing antibodies is important for mediating antibody- dependent intracellular neutralization (ADIN) and protection against papillomavirus challenge. Specific Aim 2A: To determine whether binding of the Fc of L2-specific antibodies to TRIM21 contributes to antibody-dependent intracellular neutralization and if this is restricted to particular epitopes. Specific Aim 2B: To determine the contribution of TRIM21 to protection by HPV vaccination.

描述（由申请人提供）：我们的总体目标是了解L2特异性抗体如何对HPV感染产生广泛的保护。在这里，我们剖析了为什么需要通过伽马分泌酶进行L2和内膜内切割，以及在感染过程中多个步骤中和中和的L2抗体如何通过中和。在最初的资金perio中，我们对HPV的感染绝对需要γ-分泌酶（Karanam等，2010），但只有HPV开始在晚期内部内部和伽马分泌酶的蛋白水解活性中拆卸后才需要γ-分泌酶。为了逃离与病毒基因组相关的L2晚期内体。最近的发现表明，L2在残基45-67之间具有跨膜样结构域，这对于感染至关重要。我们建议推定的跨膜区域刺穿内体膜，而γ-分泌酶的裂解释放了L2的羧基末端，将病毒基因组带到核。 L2的两个钥匙中和表位均位于L2假定的跨膜结构域的两侧。即使在病毒与细胞结合几个小时后，添加L2抗体也被中和，这表明封锁发生在感染后期。因此，我们提出：假设1：通过γ-分泌酶在其推定的跨膜区域内的L2裂解对于内体逃生至关重要，并且受到L2特异性中和抗体的抑制。特定目标1a：确定膜插入所需的L2残基，以及它们是否通过促进内体逃生来促进感染性。特定目标1B：确定HPV感染期间伽马泌物酶裂解的功能作用和部位，以及L2特异性中和抗体和伽马泌尿酶抑制剂对内体逃生的影响。通常假定抗体以纯粹的细胞外影响进行保护，因为它们通常在细胞质中没有发现。然而，最近描述了依赖于抗体的细胞内中和（ADIN）的TRIM21依赖性机制。 TRIM21是一种细胞质高亲和力FC受体，用于细胞质中病毒相关抗体的FC，从而触发抗体结合病毒的降解。我们的新发现表明，在由L2抗体介导的保护中，阿丁可能是重要的防御。假设2：L2特异性中和抗体的FC区域对于介导抗体依赖性的细胞内中和（ADIN）和防止乳头瘤病毒挑战的保护很重要。特定目标2a：确定L2特异性抗体对TRIM21的FC的结合是否有助于抗体依赖性细胞内中和，并且是否仅限于特定的表位。具体目标2B：确定TRIM21通过HPV疫苗接种保护的贡献。