Immune Induction to Genital Herpes Simplex Virus 2

描述（由申请人提供）：单纯疱疹病毒-2（HSV-2）是最常见的性传播感染（STI）之一，在美国的患病率为4500万。HSV-2通过暴露于生殖器粘膜表面传播，导致在骶神经节中建立潜伏期，从而引起不治之症。虽然HSV-2相关的症状可以用抗病毒剂暂时治疗，但没有疫苗可用于预防这种疾病。为了开发有效的HSV-2疫苗，需要清楚地了解相关粘膜部位内产生免疫应答的机制。目前，对女性生殖器粘膜内免疫应答的启动过程知之甚少。因此，拟议的研究的目标是了解诱导机制的局部保护性免疫介导的抗原呈递树突状细胞（DC）在女性生殖道感染HSV-2。在本申请中，我们以第一个资助期内的发现为基础，提出了三个不同但相互关联的目标。在第一个目标中，基于我们的初步数据表明，Toll样受体（TLR）和CD4 T的需求有助于产生CTL反应后HSV-2感染，我们建议解剖的机制，这两个信号使DC总理CTL反应对生殖器疱疹感染。接下来，基于我们的证明，TLR介导的HSV-2感染的感染的阴道角质形成细胞的识别是必要的粘膜下DC（SMDC），以诱导强大的Th1免疫在体内，在第二个目的，我们建议确定角质形成细胞产生的因素负责这种效果，并检查它们在体内抗HSV-2免疫的相关性。在最后的目的，我们建议检查在体内的重要性，SMDC在免疫启动产生SMDC诱导敲除小鼠。我们将建立一个转基因小鼠系，其中CD301b基因的启动子区，特异性表达的SMDC，将用于表达白喉毒素（DT）受体-GFP融合分子。在用DT处理此类小鼠后，将实现SMDC的选择性消耗。该目标将汇集目标1和2中的知识和进展，并提供关于上皮细胞、SMDC和其他APC在HSV-2先天识别中的相互作用的基本见解，从而导致产生适应性免疫。通过提供对阴道DCs如何协调HSV-2生殖器感染后免疫产生的基本理解，这些研究将有助于建立设计针对生殖器疱疹和其他有害STI疾病的免疫干预和预防措施的关键基础。公共卫生相关性：单纯疱疹病毒-2（HSV-2）是生殖器疱疹的病原体，是一种高度流行的性传播感染（仅在美国就有4500万人）。一旦获得，HSV-2会导致终身无法治愈的衰弱性疾病，目前还没有可用的疫苗。该资助申请旨在研究女性生殖器粘膜中局部树突状细胞启动针对HSV-2感染的免疫反应的机制-通过该项目获得的理解将有助于建立关键基础，以设计针对生殖器疱疹和其他有害性传播疾病的免疫干预和预防措施。

DESCRIPTION (provided by applicant): Herpes simplex virus-2 (HSV-2) is one of the most common sexually transmitted infections (STIs) with a prevalence of 45 million in the USA. HSV-2 is transmitted via exposure at the genital mucosal surfaces, leading to the establishment of latency in the sacral ganglia, causing an incurable disease. Although HSV-2- related symptoms can be treated temporarily with antiviral agents, there are no vaccines available for the prevention of this disease. Towards developing an effective HSV-2 vaccine, a clear understanding of the mechanism by which immune responses are generated within the relevant mucosal sites is necessary. Currently, the process of initiation of immune responses within the female genital mucosa is poorly understood. Thus, the goal of the proposed studies is to understand the inductive mechanism of local protective immunity mediated by antigen presenting dendritic cells (DCs) in the female genital tract following infection with HSV-2. In this application, we build on the discoveries made in the first funding period and propose three distinct yet inter-related Aims. In the first Aim, based on our preliminary data demonstrating the requirement for both Toll-like receptor (TLR) and CD4 T help in generating CTL responses following HSV-2 infection, we propose to dissect the mechanisms by which these two signals enable DCs to prime CTL responses against genital herpes infection. Next, based on our demonstration that TLR-mediated recognition of HSV-2 infection by the infected vaginal keratinocytes is necessary for submucosal DCs (SMDCs) to induce robust Th1 immunity in vivo, in the second Aim, we propose to identify the keratinocyte- produced factors responsible for this effect, and to examine their relevance in anti-HSV-2 immunity in vivo. In the final Aim, we propose to examine the in vivo importance of SMDCs in immune priming by generating SMDC inducible-knockout mice. We will create a transgenic mouse line in which the promoter region of the CD301b gene, specifically expressed by the SMDCs, will be used to express diphtheria toxin (DT) receptor- GFP fusion molecule. Upon treatment of such mice with DT, selective depletion of SMDCs will be achieved. This Aim will bring together the knowledge and advances made in Aims 1 & 2 and provide fundamental insights on the interplay between the epithelial cells, SMDCs and other APCs in innate recognition of HSV-2 leading to the generation of adaptive immunity. By providing basic understanding of how vaginal DCs orchestrate the generation of immunity following HSV-2 genital infection, these studies will help to establish critical foundation with which to design immunological interventions and preventative measures against genital herpes and other deleterious STI diseases. PUBLIC HEALTH RELEVANCE: Herpes simplex virus-2 (HSV-2), a causative agent of genital herpes, is a highly prevalent (45 million in the USA alone) sexually transmitted infection. Once acquired, HSV-2 causes a life-long incurable debilitating disease for which no vaccines are currently available. This grant application proposes to examine the mechanism by which local dendritic cells in the female genital mucosa initiate immune responses against HSV-2 infection - the understanding obtained through the project will help to establish critical foundation with which to design immunological interventions and preventative measures against genital herpes and other deleterious sexually transmitted diseases.