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，通常是良性病毒。然而，HCMV经常有助于拒绝移植患者的器官，并导致全身性疾病和新生儿中枢神经系统发育的缺陷。 HCMV感染许多不同的细胞类型，包括上皮和内皮细胞，神经胶质细胞，成纤维细胞和单核细胞巨噬细胞。这种广泛的热情是通过涉及不同病毒糖蛋白的不同入口途径进入不同细胞类型的能力来制备的，包括：GH/GL/GL/UL128-131，表示Pentamer，The Trimer，GH/GL/GO。我们的HCMV如何进入上皮细胞和内皮细胞的模型表明，HCMV的三聚体与细胞表面受体结合，例如PDGFRα病毒是内在化的，五聚体在内体中起作用，以促进成纤维细胞中的GB介导的融合，然后促进与细胞膜的病毒素包膜的融合。有第三种形式的GH/GL，具有GB，GB-GH/GL的GH/GL复合物，我们不知道GB-GH/GL是否在病毒进入中促进。鉴于它们在病毒进入中的重要性，触发和五聚体也是抗体（ABS）的重要靶标，被认为是HCMV疫苗设计中的关键参与者。提出了四个目标：目标1。表征HCMV进入成纤维细胞以及上皮和内皮细胞的途径，并确定触发和五聚体功能的位置。该目标将检验以下假设，即触发与细胞受体的结合导致细胞表面流量，然后将病毒颗粒内部化到细胞中，下游五聚体介导的作用促进从内体出口的病毒出口到细胞质。目标2。确定触发和触发的结构：PDGFRα，定义触发结构/功能关系并识别其他触发受体。我们的受体PDGFR具有触发的初步结构，我们将扩展这些结构研究，并使用定向的突变体来测试功能。其他研究将确定对进入上皮细胞和内皮细胞重要的触发受体。目的3。研究HCMV GB-GH/GL复合物的功能。我们将通过使用GB和GH/GL的突变体形式来阻止GB-GH/GL的组装来检验GB-GH/GL对于HCMV进入至关重要的假设。目标4。要表征人类血清中的触发特异性ABS，并与Pentamer ABS进行比较。我们对人血清中的三聚体和五聚体特异性ABS进行了惊人的观察，以中和HCMV。我们将扩展这些研究表征三聚体特异性ABS的患病率和效力，并确定这些AB识别的三聚体中的表位。