Mechanisms of Protective Local Immunity in Human Female Reproductive Tract

描述(由申请人提供)：性传播疾病(STDS)的流行在全球范围内有所增加，造成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)的疫苗开发，如艾滋病毒和单纯疱疹病毒2型(HSV-2)，基本上没有成功。阴道和宫颈外的粘膜表面既是入口点，也是防止性传播病原体的屏障。组织定位的记忆T细胞通过直接针对感染细胞和从循环中招募记忆T细胞，在皮肤和粘膜中提供了第一道防线。研究表明，CD8T细胞不仅可以渗透到感染部位，而且可以在生殖器皮肤和粘膜中持续存在。 病毒清除后的时间段。我们最近发表的工作(《自然》，2013)表明，HSV-2生殖器病变愈合后，驻留在人生殖器皮肤中的CD8T细胞表达CD8？？同源二聚体作为辅助受体，而不是异二聚体CD8β，后者主宰血液循环中的CD8T细胞。CD8的表面表达？？同源二聚体也是肠道上皮细胞上皮内淋巴细胞(IEL)和粘膜相关不变T细胞(MAIT)的特征。CD8？？已被提议保存高亲和力效应T细胞以用于长期的粘膜记忆。是否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，我们建议研究CD8？？的免疫学相关性。通过外阴、阴道和宫颈外组织的活检，对人类女性生殖道(FRT)中的T细胞以及CD4T细胞的驻留状态进行了研究。这项建议的总体目标是确定记忆CD8和CD4T细胞在FRT组织间隔中的驻留状态，并确定其局部保留、功能和自我更新能力的机制。利用特定细胞类型的激光捕获显微切割(LCM)、转录图谱、高通量测序技术和多色共聚焦显微镜相结合的方法，我们的目标是：1)定义CD_4、CD_8、CD_8的空间动力学；和CD8？？T细胞在外阴、阴道和宫颈中的分布；2)决定驻留在人类FRT中的记忆T细胞的组织保留和功能的机制；以及3)确定促进驻留T细胞局部增殖的微环境信号。我们相信，对人类FRT中细胞免疫的彻底研究将扩大我们对STI获得和传播的解剖部位组织驻留免疫的了解。然后，这些研究可以直接为未来疫苗和免疫治疗方法的设计提供信息，这些方法可以诱导组织常驻免疫反应，希望有效地防止性传播感染。

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.