Herpes Simplex Virus Entry Receptors in Immune Memory and Newborn Infection

描述(由申请人提供)：目标：此申请定义了一个计划，以促进有前途的初级调查员在指导环境下的研究生涯。成功完成研究将使研究人员开始作为一名独立的临床科学家的职业生涯，进行旨在创造和评估单纯疱疹病毒(HSV)治疗方法和候选疫苗的转译研究，相关目标是提高对HSV免疫的了解。这一应用的具体项目调查了HSV与其主要进入受体在新生儿疾病中相互作用的重要性，以及HSV糖蛋白和免疫信号蛋白之间的相互作用对HSV记忆免疫的影响。背景：单纯疱疹病毒是人类常见的病原体，感染60%以上的美国成年人，新生儿尤其容易患上严重疾病。单纯疱疹病毒最初是在与两个主要受体之一--HVEM和Nectin相互作用后感染细胞的。先前发表的数据表明，神经系统疾病在很大程度上与使用Nectin的病毒进入有关；我们的初步数据表明，病毒可能通过参与Hvem来操纵记忆免疫。研究设计和方法：在本项目的特定目标1中，我们将用减毒的HSV-2经阴道感染成年雌性小鼠，用野生型病毒或改变其与HSV结合的病毒来激发对HSV的免疫。然后，我们将严格研究野生型或突变型病毒攻击后记忆淋巴细胞的反应，解决这一假设，即粘膜上的调节性T细胞反应在记忆阶段通过病毒与HIV的初始相互作用而改变。在目标2中，我们将测试HVEM参与对新生儿HSV疾病的影响。我们将使用与目标1类似的免疫学方法来测试新生小鼠的免疫力是否因接触HVEM而改变。感染途径将与人类新生儿疾病相关。在目标3中，我们将详细调查我们的初步观察结果，即HVEM不足以在新生小鼠中引起HSV疾病，观察疾病在缺乏一个或两个主要受体的小鼠中的传播和发病机制。我们还将测量不同受体在出生后早期发育不同组织中的相对表达。研究环境：应聘者建议在培养初级调查人员的职业生涯方面取得成功的环境下开发该项目。在该领域专家的监督下，该项目将为候选人的背景增加新的专业知识，包括发展小鼠新生儿感染模型，调查新生儿免疫反应，以及免疫组织化学方法。该提案中的职业发展活动包括分子生物学和免疫学的教学课程，职业咨询委员会的定期评估，以及负责任的研究行为培训。 公共卫生相关性：了解对单纯疱疹病毒(HSV)的记忆免疫有助于开发有效防止慢性感染者脱皮的疫苗，并可能导致开发治疗方法，以最大限度地减少或防止新生儿在分娩期间暴露。更好地了解不同的HSV受体在新生儿疾病发病机制中的影响，可能会改进这一人群的治疗方法。

DESCRIPTION (provided by applicant): Objectives: This application defines a program to further the research career of a promising junior investigator within a mentored setting. Successful completion would allow the investigator to initiate a career as an independent clinician-scientist, conducting translational research directed at creating and evaluating therapeutics and vaccine candidates for herpes simplex virus (HSV), with the related goal of improving the understanding of immunity to HSV. The specific project for this application investigates the importance of HSV interactions with its principal entry receptors on neonatal disease and the influence of an interaction between an HSV glycoprotein and an immune signaling protein on memory immunity to HSV. Background: Herpes simplex viruses are common human pathogens, infecting more than 60% of American adults, with newborns particularly susceptible to severe disease. HSV initially infects cells after interacting with one of two principal receptors, HVEM and nectin. Prior published data suggest that neurologic disease is largely related to viral entry using nectin; our preliminary data suggest that the virus may manipulate memory immunity by engagement of HVEM. Research design and methods: In Specific Aim 1 of this project, we will prime immunity to HSV in adult female mice by intravaginal infection with attenuated HSV-2, using either wild-type virus or virus altered to abrogate binding to HVEM. We will then rigorously investigate the memory lymphocyte response after challenge with either wild-type or mutant virus, addressing the hypothesis that regulatory T-cell responses at the mucosa are altered in the memory phase by initial viral interaction with HVEM. In Aim 2, we will test the influence of HVEM engagement on neonatal HSV disease. We will use similar immunologic methods as in Aim 1 to test whether immunity in newborn mice is altered by engagement of HVEM. Routes of infection will be relevant to neonatal disease in humans. In Aim 3, we will investigate in detail our preliminary observation that HVEM is not sufficient to cause HSV disease in newborn mice, looking at spread and pathogenesis of disease in mice lacking one or both principal receptors. We will also measure the relative expression of different receptors in different tissues during early postnatal development. Research environment: The candidate proposes to develop this project within an environment with established success at nurturing the careers of junior investigators. Under the supervision of experts in the field, this project will add new expertise to the candidate's background, including development of murine neonatal infection models, investigation of newborn immune responses, and immunohistochemical methods. Career development activities within the proposal include didactic coursework in molecular biology and immunology, regular evaluations by a career advisory committee, and training in the responsible conduct of research. Public health relevance: The understanding of memory immunity to herpes simplex virus (HSV) has implications for the development of vaccines effective at preventing shedding in chronically infected individuals, and may lead to development of therapeutic methods to minimize or prevent neonatal exposure during delivery. A better understanding of the influence of different HSV receptors on pathogenesis of neonatal disease may lead to improved therapeutics in this population.