Project 2: Virologic determinants of cCMV

项目 2：cCMV 的病毒学决定因素

• 批准号: 10215785
• 负责人: Sallie R. Permar
• 金额: $ 0.14万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-24 至 2020-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10215785
• 关键词: Active Immunization; Address; Adult; Affinity; Amniotic Fluid; Anatomy; Animal Model; Animals; Antibodies; Antibody Therapy; Bacterial Artificial Chromosomes; Blood; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; Cells; Clinical Research; Complex; Cytomegalovirus; Cytomegalovirus Infections; Cytomegalovirus Vaccines; Data; Development; Disease; Fc Receptor; Female; Fetus; Genes; Genome; Glycoproteins; Goals; Human; Immune; Immune system; Immunoglobulin G; Immunologics; Individual; Information Dissemination; Intravenous; Length; Macaca mulatta; Maternal-Fetal Transmission; Modeling; Molecular Cloning; Mutate; Mutation; Newborn Infant; Passive Immunization; Population; Preventive measure; Primates; Proteins; RL13; Reporting; Rhesus; Role; Sequence Homology; Services; Spontaneous abortion; Testing; Therapeutic; Therapeutic Intervention; Time; Tissues; Tropism; Viral; Viral Antibodies; Viral Genes; Viral Proteins; Viremia; Virus; abortion; anti-viral efficacy; base; congenital cytomegalovirus; congenital infection; design; experimental study; fetal; fetal infection; genetic manipulation; improved; in vivo; male; member; nonhuman primate; novel strategies; pregnant; programs; prophylactic; receptor; recombinant virus; reconstitution; seropositive; therapy development; tissue culture; transmission process; vaccine development; viral transmission; virology; virus genetics

ABSTRACT – Project 2: Viral determinants of fetal RhCMV transmission Human cytomegalovirus (HCMV) is the leading infectious cause of congenital disease in newborns, and the development of treatments and preventative measures requires a better understanding of the viral determinants enabling fetal transmission in CMV-naïve and CMV-immune individuals. Since HCMV is highly species-restricted, animal models of congenital infection by the corresponding animal CMVs are used to study congenital infection. Recent advances in non-human primate models for the first time enable the study of fetal transmission in a host that is anatomically very similar to humans by a virus that is highly homologous to HCMV. The goal of this project is therefore to use this model to identify and characterize viral determinants facilitating fetal transmission to ultimately support the development of specific countermeasures. Specifically, we will examine whether fetal transmission correlates with viremia and viral dissemination in the pregnant female. We will further examine the role of viral cell tropism in fetal infection and examine the hypothesis that the efficacy of antiviral antibodies is limited by viral Fc receptors. To enable the genetic manipulation of RhCMV that is required to address these questions we generated a bacterial artificial chromosome (BAC) clone representative of a low passage isolate and demonstrated fetal transmission of BAC-derived RhCMV. Using this BAC we will take advantage of our observation that RhCMV lacking the immunodominant major tegument protein pp65 displayed dramatically increased replication and dissemination in RhCMV-naïve animals to examine the role of viremia and viral dissemination in fetal transmission (Aim 1). We will further experimentally address the role of the cell tropism-determining pentameric glycoprotein complex gH/gL/UL128/UL130/UL131A in maternal/fetal transmission (Aim 2). The pentameric complex is currently a major target for vaccine development, but preliminary data unexpectedly revealed increased transmission of virus lacking this complex. Finally, we will determine whether viral Fc-Receptors limit the efficacy of hyperimmunoglobulin treatment to limit viral dissemination (Aim 3). This project will both benefit from and impact the studies described in Project 1 and require the services of all four cores. We anticipate that this project will impact the design of prophylactic and therapeutic CMV vaccines and that it will reveal novel approaches to improve antibody-based therapy.

摘要 - 项目2：胎儿RHCMV传播的病毒决定剂 人类巨细胞病毒（HCMV）是新生儿先天性疾病的主要感染原因， 制定治疗和预防措施需要更好地了解病毒 决定因素，可以在CMV-NOVING和CMV免疫个体中传播胎儿传播。由于HCMV高度高 相应动物CMV限制的物种，先天性感染的动物模型用于研究 先天性感染。非人类灵长类动物模型的最新进展首次使胎儿研究 通过与人类非常相似的宿主传播，该病毒与人类高度同源 HCMV。因此，该项目的目的是使用该模型来识别和表征病毒决定者 促进胎儿传播，最终支持特定对策的发展。具体来说， 我们将检查胎儿传播是否与孕妇的病毒血症和病毒传播相关 女性。我们将进一步研究病毒细胞疗法在胎儿感染中的作用，并研究以下假设。 抗病毒抗体的效率受病毒FC受体的限制。为了实现遗传操作 解决这些问题所需的RHCMV我们产生了人造染色体（BAC）的细菌 克隆代表低通道分离株，并显示出BAC衍生的RHCMV的胎儿传播。 使用此BAC，我们将利用我们的观察，即RHCMV缺乏免疫主导专业 Tegument蛋白PP65在RHCMV-NOï中显示出显着增加的复制和传播 动物检查病毒血症和病毒传播在胎儿传播中的作用（AIM 1）。我们将进一步 通过实验解决细胞对端主的五聚糖蛋白复合蛋白复合物的作用 母体/胎儿传播中的GH/GL/GL/UL128/UL130/UL131A（AIM 2）。五聚会综合体当前是 疫苗开发的主要目标，但初步数据出乎意料地发现 缺乏这种复合物的病毒。最后，我们将确定病毒FC受体是否限制 高免疫球蛋白治疗以限制病毒传播（AIM 3）。这个项目将从中受益，并 影响项目1中描述的研究，并需要所有四个核心的服务。我们预料到了 项目将影响预防性和治疗性CMV疫苗的设计，并将揭示新颖 改善基于抗体的治疗的方法。