Human Cytomegalovirus Entry into Cells Mediated by Pentamer and Trimer Complexes

五聚体和三聚体复合物介导的人巨细胞病毒进入细胞

基本信息

批准号：
10687819
负责人：
Theodore S Jardetzky
金额：
$ 73.27万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-17 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10687819
关键词：
Address Affect Antibodies Antibody Response Benign Binding Biochemical Blood Vessels Capsid Cell Surface Receptors Cell surface Cells Cellular Membrane Complex Cytomegalovirus Cytomegalovirus Vaccines Cytoplasm Data Defect Development Disease Endosomes Endothelial Cells Endothelium Epithelial Cells Epithelium Epitopes Fetal Development Fibroblasts Foundations Glycoproteins Grant Hepatitis Hepatocyte Herpesviridae Human Image Immune Targeting Immune system Immunity Immunize Infection Macrophage Maps Mediating Membrane Fusion Modeling Molecular Conformation Mutagenesis Nervous System Neuroglia Newborn Infant Oryctolagus cuniculus PDGFRB gene Pathology Pathway interactions Peptides Platelet-Derived Growth Factor alpha Receptor Pneumonia Prevalence Production Property Proteins Receptor Cell Role Site Structure Structure-Activity Relationship Systemic disease Testing Transplant Recipients Tropism Vaccines Viral Viral Fusion Proteins Virion Virus Virus Diseases cell type design experimental study follow-up graft failure human imaging immunosuppressed in vivo interest monocyte mutant neonate neutralizing antibody organ transplant rejection particle receptor synergism trafficking

项目摘要

Human cytomegalovirus (HCMV) is a ubiquitous, usually benign virus. Nevertheless, HCMV frequently contributes to rejection of organs in transplant patients and causes systemic disease and defects in the development of the CNS in neonates. HCMV infects many different cell types including epithelial and endothelial, glial cells, fibroblasts and monocyte-macrophages. This broad tropism is facilitated by a capacity to enter different cell types via distinct entry pathways involving different viral glycoproteins including: gH/gL/UL128-131, denoted the pentamer, gH/gL/gO, the trimer. Our model for how HCMV enters epithelial and endothelial cells suggests that HCMV trimers bind to cell surface receptors, e.g. PDGFRα viruses are internalized and pentamer acts in endosomes to promote gB-mediated fusion in fibroblasts, then of the virion envelope with cellular membranes. There is a third form of gH/gL, a complex of gH/gL with gB, gB-gH/gL and we do not know whether gB-gH/gL promotes in virus entry. Given their importance in virus entry, trimer and pentamer are also important targets of antibodies (Abs) and are considered key players in the design of HCMV vaccines. Four aims are proposed: Aim 1. To characterize pathways of HCMV entry into fibroblasts and epithelial and endothelial cells and determine where trimer and pentamer function. This aim will test the hypothesis that trimer binding to cellular receptors leads to cell surface traffic followed by internalization of virus particles into cells and downstream pentamer-mediated effects promoting virus exit from endosomes into the cytoplasm. Aim 2. To determine the structures of trimer and trimer:PDGFRα, define trimer structure/function relationships and identify other trimer receptors. We have a preliminary structure of trimer with its receptor PDGFR We will extend these structural studies and use site directed mutants to test function. Other studies will identify trimer receptors important for entry into epithelial and endothelial cells. Aim 3. To investigate how HCMV gB-gH/gL complexes function. We will test the hypothesis that gB- gH/gL is important for HCMV entry by using mutant forms of gB and gH/gL to block assembly of gB-gH/gL. Aim 4. To characterize trimer- specific Abs in human sera and compare to pentamer Abs. We made striking observations that trimer- and pentamer-specific Abs in human sera synergize to neutralize HCMV. We will extend these studies characterize the prevalence and potency of trimer-specific Abs and identify the epitopes in trimer recognized by these Ab.

人类巨细胞病毒 (HCMV) 是一种普遍存在的、通常是良性的病毒，然而，HCMV 经常导致移植患者的器官排斥，并导致新生儿中枢神经系统发育缺陷。、神经胶质细胞、成纤维细胞和单核细胞巨噬细胞通过不同的进入进入不同细胞类型的能力促进了这种广泛的趋向性。涉及不同病毒糖蛋白的途径包括：gH/gL/UL128-131（表示五聚体）、gH/gL/gO（表示三聚体）。我们关于 HCMV 如何进入上皮和内皮细胞的模型表明，HCMV 三聚体与细胞表面受体（例如 PDGFRα）结合。病毒被内化，五聚体在内体中发挥作用，促进成纤维细胞中 gB 介导的融合，然后gH/gL 有第三种形式，即 gH/gL 与 gB 的复合物，即 gB-gH/gL，鉴于 gB-gH/gL 的重要性，我们不知道 gB-gH/gL 是否会促进病毒进入。病毒进入、三聚体和五聚体也是抗体 (Abs) 的重要靶点，被认为是 HCMV 疫苗设计中的关键因素，提出了四个目标：目标 1. 描述 HCMV 进入的途径。成纤维细胞、上皮细胞和内皮细胞，并确定三聚体和五聚体在何处发挥作用，该目标将检验以下假设：三聚体与细胞受体的结合导致细胞表面运输，然后病毒颗粒内化到细胞中，并促进下游五聚体介导的病毒从内体退出的作用。目标 2. 确定三聚体和三聚体：PDGFRα 的结构，定义三聚体结构/功能关系并鉴定其他三聚体受体。三聚体及其受体 PDGFR 我们将扩展这些结构研究并使用定点突变体来测试功能。目标 3. 研究 HCMV gB-gH/gL 复合物的功能。我们将通过使用 gB 和 gH/gL 的突变形式来阻止 gB-gH/gL 的组装来检验 gB-gH/gL 对于 HCMV 进入很重要的假设。目标 4. 表征人类血清中的三聚体特异性抗体并与五聚体抗体进行比较我们发现人类血清中的三聚体和五聚体特异性抗体可协同中和 HCMV 我们将扩展这些表征三聚体的流行性和效力的研究。 -特异性抗体并鉴定这些抗体识别的三聚体中的表位。