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 传播的病毒决定因素 人类巨细胞病毒 (HCMV) 是新生儿先天性疾病的主要感染原因， 治疗和预防措施的开发需要更好地了解病毒 由于 HCMV 具有高度传染性，因此在未接触过 CMV 的个体和对 CMV 免疫的个体中能够进行胎儿传播的决定因素。 使用相应动物 CMV 先天性感染的物种限制的动物模型来研究 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成为可能。 通过与人类高度同源的病毒在解剖学上与人类非常相似的宿主中传播 因此，该项目的目标是使用该模型来识别和表征病毒决定因素。 促进胎儿传播，最终支持制定具体对策。 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 我们将进一步研究病毒细胞趋向性在胎儿感染中的作用，并检验以下假设： 抗病毒抗体的功效受到病毒 Fc 受体的限制，以实现基因操作。 解决这些问题所需的 RhCMV 我们生成了细菌人工染色体 (BAC) 代表低传代分离株的克隆，并证明了 BAC 衍生的 RhCMV 的胎儿传播。 使用此 BAC，我们将利用我们的观察结果，即 RhCMV 缺乏免疫显性主要 外皮蛋白 pp65 在未经 RhCMV 处理的细胞中表现出显着增加的复制和传播 我们将进一步研究病毒血症和病毒传播在胎儿传播中的作用（目标 1）。 通过实验解决细胞趋向性决定五聚体糖蛋白复合物的作用 gH/gL/UL128/UL130/UL131A 在母​​体/胎儿传播中的作用（目标 2）。 疫苗开发的主要目标，但初步数据意外显示传播增加 最后，我们将确定病毒 Fc 受体是否会限制病毒的功效。 限制病毒传播的高免疫球蛋白治疗（目标 3）将受益于该项目。 影响项目 1 中描述的研究并需要所有四个核心的服务，我们预计这会发生。 该项目将影响预防性和治疗性巨细胞病毒疫苗的设计，并将揭示新的 改善基于抗体的治疗的方法。