Role of epididymal dendritic cells in male reproductive function

附睾树突状细胞在男性生殖功能中的作用

• 批准号: 8158869
• 负责人: Nicolas Da Silva
• 金额: $ 37.34万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-12 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8158869
• 关键词: Affect; Antigens; Autoimmune Responses; Autoimmunity; Behavior; Biological Assay; Blood; CD8B1 gene; CX3CL1 gene; Cell Line; Characteristics; Data; Dendritic Cells; Development; Disease; Distal; Duct (organ) structure; Environment; Epididymis; Epithelial Cells; Epithelium; Equilibrium; Escherichia coli; Fertility; Flow Cytometry; Fluorescence; Goals; ITGAX gene; Image; Imaging Techniques; Immune Tolerance; Immune response; Immune system; Impairment; In Vitro; Infection; Infertility; Inflammation; Injection of therapeutic agent; Intervention; Label; Laboratory Study; Lead; Ligation; Male Infertility; Medical; Microscopy; Morphology; Mucous Membrane; Mus; Nature; Phenotype; Play; Population; Positioning Attribute; Process; Puberty; Public Health; Relative (related person); Reproductive Physiology; Resolution; Role; Sampling; Site; Specificity; Sperm Maturation; Spermatogenic Cell; T-Lymphocyte; Tail; Testing; Testis; Texas red; Time; Transgenic Mice; Tube; Vasectomy; Vasovasostomy; Work; base; cancer cell; comparative; epididymis head; immune activation; in vivo; intraepithelial; male; microorganism; particle; pathogen; postnatal; prevent; reproductive; reproductive function; sperm cell

DESCRIPTION (provided by applicant): Infection or inflammation of the male reproductive tract often lead to permanent infertility. Yet, one of the most understudied aspects of male reproductive physiology is the ability of the excurrent duct to protect maturing spermatozoa against an autoimmune response, while initiating very efficient immune activation against pathogens. The present application is based on our new discovery that a dense dendritic cell network populates the epididymis. We showed that epididymal dendritic cells (eDCs) establish intimate interactions with the epididymal mucosa and can send dendritic projections between epithelial cells toward the lumen. eDCs have the ability to present antigens to CD4+ and CD8+ T cels in vitro. We hypothesize that eDCs can interact with the luminal content, including spermatozoa, and regulate the balance between immune tolerance and inflammation. Aim 1a wil characterize eDCs using high resolution imaging in CD11c-EYFP and CX3CR1-GFP transgenic mice, comparative flow cytometry analysis and antigen-presenting assays before, during and after puberty. We postulate that the eDC network is reorganized during puberty, when sperm begin populating the excurrent duct. Aim 1b wil examine the regional specificities of eDC subsets in the context of the dual function of the epididymis. eDCs will be isolated from the proximal epididymal region (primarily involved in sperm maturation and protection against an autoimmune response), and distal region (more susceptible to infections by ascending pathogens). Comparative flow cytometry analysis and antigen-presenting capability will be determined in proximal and distal eDC populations. Aim 2 wil determine the role of eDCs in the homeostatic and inflamed mouse epididymis, using conventional fluorescence and intravital multiphoton microscopy, and flow cytometry analysis. Aim 2a wil test the hypothesis that eDCs have the ability to sample and process luminal antigens including those present in sperm. Aim 2b will analyze the behavior of eDCs in the epididymis after injection with LPS (to mimic infections), vasectomy (downstream blockade of luminal flow) or efferent duct ligation (upstream blockade of luminal flow). The goal of this application is to characterize the nature and functions of the eDC network to provide new frameworks for the treatment of male immunological infertility (which is a major public health issue) and, ultimately, for the control of male fertility. PUBLIC HEALTH RELEVANCE: Spermatozoa acquire their fertilizing capacity during their transit through the epididymis, a long tube located downstream of the testis. Our laboratory studies how dendritic cells, which are among the most potent regulators of the immune system, work together with other cells lining the epididymal tube to protect spermatozoa against pathogens and autoimmunity. These results will provide new frameworks for the treatment of immunological male infertility.

描述(申请人提供)：男性生殖道感染或发炎通常会导致永久性不育。然而，男性生殖生理学最不被研究的方面之一是流出管保护成熟精子免受自身免疫反应的能力，同时启动非常有效的针对病原体的免疫激活。目前的应用是基于我们的新发现，即致密的树突状细胞网络分布在附睾部。我们发现，附睾树突状细胞(EDCs)与附睾粘膜建立了密切的相互作用，并能在上皮细胞之间向管腔发出树突状投射。在体外，EDCs具有向CD4+和CD8+T细胞递呈抗原的能力。我们假设EDCs可以与包括精子在内的腔内容物相互作用，并调节免疫耐受和炎症之间的平衡。目的研究CD11c-EYFP和CX3CR1-GFP转基因小鼠在青春期前、青春期和青春期后的高分辨率成像、比较流式细胞仪分析和抗原提呈实验。我们推测，在青春期，当精子开始填充流出管道时，EDC网络发生了重组。目的1b将在附精的双重功能的背景下检查EDC亚群的区域特异性。EDCs将从附睾部近端(主要参与精子成熟和抵御自身免疫反应)和远端(更容易受到上升病原体的感染)分离。比较流式细胞术分析和抗原提呈能力将在近端和远端EDC人群中确定。目的2应用常规荧光显微镜、活体多光子显微镜和流式细胞术分析，探讨内皮细胞在动态平衡和炎症过程中的作用。目的2a将检验EDCs具有采集和处理管腔抗原的能力的假设，包括存在于精子中的那些。目的2b将分析注射脂多糖(模拟感染)、输精管结扎术(阻断管腔下游血流)或输精管结扎术(阻断管腔血流上游)后附睾内皮细胞的行为。这项应用的目的是确定EDC网络的性质和功能，以便为治疗男性免疫性不育(这是一个重大的公共卫生问题)并最终控制男性生育能力提供新的框架。 与公共卫生相关：精子在通过附睾处获得受精能力，附睾处是位于睾丸下游的一根长管。我们的实验室研究了树突状细胞是如何与附睾管内的其他细胞合作保护精子免受病原体和自身免疫的影响。树突状细胞是免疫系统最有效的调节细胞之一。这些结果将为免疫性男性不育的治疗提供新的框架。