Placental trophoblast infection and TLR mediated response to congenital CMV

描述（由申请方提供）：开发针对先天性巨细胞病毒（CMV）的有效干预策略是一项主要的公共卫生优先事项。然而，候选CMV疫苗策略在临床前动物模型中完全预防先天性感染方面并不成功，并且有效的疫苗仍然是一个难以捉摸的目标。CMV经胎盘感染的致病性定义不明确，更好的理解可能为开发新的干预措施或疫苗策略提供新的见解。在这个提议中，我们将研究胎盘感染的两个关键方面：（1）Toll样受体（TLR）介导的胎盘对CMV的先天免疫反应;（2）病毒嗜性和胎盘感染。通过在关键胎盘细胞（滋养层细胞）上表达的TLR介导的炎性先天免疫应答是控制细菌和病毒的胎盘侵入先天性感染的潜在重要方面。已知CMV被多种TLR识别，但胎盘TLR介导的对CMV的应答定义不清，CMV潜在地绕过胎盘先天免疫并感染子宫内胎儿的机制也是如此。我们假设成功的先天性感染依赖于CMV的能力， 有效地进入和篡夺表达TLR的滋养层细胞，滋养层细胞是胎盘屏障和胎盘先天免疫的重要组成部分。新发现的病毒进入细胞的内吞途径可能是胎盘侵入和上皮滋养层细胞感染的重要因素。我们建议确定TLR介导的先天性免疫反应的胎盘CMV感染的意义，在唯一的小动物模型（豚鼠）先天性CMV。此外，使用相同的动物模型，我们将确定重要的嗜性基因，这些基因可能使病毒能够在胎盘中建立感染灶并随后感染子宫内的胎儿。这些研究将采用传统的组织病理学和免疫组织化学方法以及新技术，包括在动物模型中使用病毒传播的生物发光成像。

DESCRIPTION (provided by applicant): Development of an effective intervention strategy against congenital cytomegalovirus (CMV) is a major public health priority. However, candidate CMV vaccines strategies have been unsuccessful in completely preventing congenital infection in a pre-clinical animal model and an effective vaccine remains an elusive goal. The pathogenicity of CMV transplacental infection is poorly defined and a better understanding may provide fresh insight for the development of a novel intervention or vaccine strategy. In this proposal, we will examine two key aspects of placental infection: (1) Toll-like receptor (TLR) mediated innate immune response of the placenta to CMV; (2) Viral tropism and placental infection. The inflammatory innate immune response mediated via TLRs expressed on key placental cells (trophoblasts) is potentially an important aspect of controlling placental invasion congenital infection by both bacteria and viruses. CMV is known to be recognized by multiple TLRs but the placental TLR mediated response to CMV is poorly defined as is the mechanism by which CMV potentially circumvents the placental innate immunity and infects the fetus in utero. It is our hypothesis that successful congenital infection is reliant upon the ability of CMV to efficiently enter and usurp TLR expressing trophoblast cells, an important component of the placental barrier and placental innate immunity. Potentially, the newly identified endocytic pathway of viral cell entry could be an important factor in the invasion of the placenta and infection of epithelial trophoblasts. We propose to determine the significance of the TLR mediated innate immune response of the placenta to CMV infection in the only small animal model (guinea pig) for congenital CMV. Additionally, using the same animal model, we will define important tropism genes that potentially enable the virus to establish infectious foci in th placenta and subsequently infect the fetus in utero. These studies will employ conventional histopathological and immunohistochemical approaches as well as novel techniques including the use of bioluminescence imaging of viral dissemination in the animal model.