Mechanisms of Protective Local Immunity in Human Female Reproductive Tract

描述（由申请人提供）：性传播疾病（STD）的流行率在全球范围内增加，并带来严重的医疗和心理后果。尽管做出了重大努力，但旨在预防或治疗性传播感染（STI），如艾滋病毒和单纯疱疹病毒2型（HSV-2）的疫苗开发在很大程度上并不成功。阴道和外宫颈的粘膜表面既是进入点，也是防止性传播病原体的屏障。组织定位的记忆T细胞通过直接靶向受感染的细胞以及通过从循环中募集记忆T细胞来提供皮肤和粘膜中的第一道防线。我们已经证明，CD 8 T细胞不仅浸润到感染部位，而且在生殖器皮肤和粘膜中持续存在很长时间。 病毒清除后的时间段。我们最近发表的工作（Nature，2013）表明，HSV-2生殖器病变愈合后，驻留在人类生殖器皮肤中的CD 8 T细胞表达CD 8？同源二聚体作为共受体，而不是异源二聚体CD 8？？，其支配血液循环的CD 8 T细胞。CD8？？同源二聚体也是肠上皮中的上皮内淋巴细胞（IEL）和粘膜相关不变T细胞（MAIT）的特征。CD8？？已经提出保存高亲和力效应T细胞用于长期粘膜记忆。CD8？表达是人类外周组织驻留记忆的一般机制，目前尚不清楚。在这里，我们建议调查的免疫相关性CD 8？？人类女性生殖道（FRT）中的T细胞，使用外阴、阴道和宫颈外组织活检，以及CD 4 T细胞的驻留状态。该提案的总体目标是定义记忆性CD 8和CD 4 T细胞在FRT组织区室中的驻留状态，并确定强调其局部保留、功能和自我更新能力的机制。利用细胞类型特异性激光捕获显微切割（LCM）、转录谱分析、高通量测序技术和激光共聚焦显微镜技术相结合的方法，我们的目标是：1）确定CD 4、CD 8？CD8？外阴、阴道和宫颈中的T细胞; 2）确定驻留在人FRT中的记忆T细胞的组织保留和功能的机制;和3）定义促进驻留T细胞的局部增殖的微环境线索。我们相信，在人类FRT细胞免疫的彻底调查，将扩大我们的知识，组织常驻免疫的STI收购和传播的解剖部位。然后，这些研究可以直接为未来疫苗和免疫方法的设计提供信息，这些疫苗和免疫方法可以引发组织驻留免疫应答，以期有效地预防性传播感染。

DESCRIPTION (provided by applicant): The prevalence of sexually transmitted diseases (STDs) has increased globally with severe medical and psychological consequences. Despite major efforts, vaccine development aiming to either prevent or treat sexually transmitted infections (STIs), such as HIV and herpes simplex virus type 2 (HSV-2), has been largely unsuccessful. The mucosal surface of the vagina and ectocervix serves as both the point of entry as well as a barrier against sexually transmitted pathogens. Tissue-localized memory T cells provide a first line of defense in the skin and mucosa, by targeting infected cells directly and also by recruiting memory T cells from the circulation. We have shown that CD8 T cells not only infiltrate to the site of infection, but also persist in genital skin and mucosa for prolonged time periods after viral clearance. Our recently published work (Nature, 2013) indicates that CD8 T cells resident in human genital skin post healing of HSV-2 genital lesions express the CD8?? homodimer as a co-receptor instead of the heterodimeric CD8??, which dominates blood circulating CD8 T cells. Surface expression of the CD8?? homodimer is also characteristic of intraepithelial lymphocytes (IEL) resident in the gut epithelium and mucosal associated invariant T cells (MAIT). CD8?? has been proposed to preserve high-affinity effector T cells for long-lived mucosal memory. Whether CD8?? expression is a general mechanism for tissue resident memory in the human periphery is currently unclear. Here, we propose to investigate the immunological relevance of CD8?? T cells in the human female reproductive tract (FRT) using biopsies of vulva, vagina and ectocervical tissue, as well as the residency status of CD4 T cells. The overall goal of this proposal is to define the resident statuses of memory CD8 and CD4 T cells in FRT tissue compartments and to determine the mechanisms underlining their local retention, function and self- renewal capability. Using a combined approach of cell-type-specific laser capture microdissection (LCM), transcriptional profiling, high-throughput sequencing technology and multicolor confocal microscopy, we aim to 1) define the spatial dynamics of CD4, CD8?? and CD8?? T cells in the vulva, vagina and cervix; 2) determine the mechanism of tissue retention and function of memory T cells resident in human FRT; and 3) define microenvironmental cues promoting local proliferation of resident T cells. We believe a thorough investigation of cellular immunity in human FRT will broaden our knowledge of tissue resident immunity at the anatomical site of STI acquisition and transmission. These studies can then directly inform design of future vaccines and immunotherapeutic approaches that elicit tissue resident immune response with the hope of effectively protecting against STIs.