Proton-secreting epithelial cells as key modulators of epididymal mucosal immunity - Administrative Supplement

质子分泌上皮细胞作为附睾粘膜免疫的关键调节剂 - 行政补充

• 批准号: 10833895
• 负责人: Maria Agustina Battistone
• 金额: $ 11.49万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10833895
• 关键词: 3-Dimensional; Ablation; Address; Administrative Supplement; Antigens; Autoimmunity; Bacterial Antigens; Bioinformatics; Cell Communication; Cell Separation; Cells; Clear Cell; Communication; Confocal Microscopy; Data; Development; Diphtheria Toxin; Disease; Environment; Epididymis; Epididymitis; Epithelial Cells; Epithelium; Equilibrium; Flow Cytometry; Goals; Health; Immune; Immune Tolerance; Immune response; Immunology; Infection; Inflammation; Injections; Injury; Knowledge; Laboratory Study; Male Contraceptive Agents; Male Infertility; Mediator; Mission; Mononuclear; Mucosal Immunity; Mucous Membrane; Phagocytes; Play; Positioning Attribute; Protons; Public Health; Regulatory T-Lymphocyte; Reproductive Biology; Reproductive Health; Reproductive Physiology; Research; Role; Specialized Epithelial Cell; Sperm Maturation; Spermatic Cord Torsion; Sterility; Surveys; Techniques; Testis; Transgenic Mice; Tube; United States National Institutes of Health; Work; cell motility; cell type; human male; immune activation; immunoregulation; innovation; insight; interdisciplinary approach; male; mouse model; novel; pathogen; prevent; sperm cell; transcriptomics

PROJECT SUMMARY/ABSTRACT One of the most intriguing and understudied aspects of male reproductive physiology is the ability of the epididymis to prevent the development of immune responses against autoantigenic spermatozoa, while initiating very efficient immune responses against pathogens. The long-term goal of this application is to develop new strategic therapies for common disorders such as male infertility and epididymitis, and to identify novel targets for male-contraception. The overall objective is to elucidate how epididymal epithelial clear cells (CCs) communicate with immunocytes to contribute to the “immune privileged” environment that is optimal for sperm maturation and storage. We have recently showed a completely novel role for CCs in immune activation and sperm tolerance. Importantly, these cells establish intimate contact with region-specific heterogeneous subsets of mononuclear phagocytes (MPs) in the epididymis. Our central hypothesis is that CCs are strategically positioned to work in a concerted manner with immune cells to survey the epididymis barrier and regulate the balance between inflammation and immune tolerance in the post-testicular environment. The rationale is based on our preliminary data showing that CCs and MPs communicate in the steady state epididymis and also in the presence of infection or injury, and that both cell types express different tolerogenic mediators allowing the modulation of sperm tolerance. Therefore, these cells play a central role in the epididymal mucosa. In the first specific aim, we will determine CC – MP communication networks that initiate immune activation during epididymitis. In this aim, two mouse models of epididymitis will be used: injection of bacterial antigens into the cauda lumen, and epididymal-testicular torsion that induces sterile inflammation. In the second specific aim, we will evaluate CC - MP crosstalk after ablation of sperm tolerance in the epididymis, induced by depletion of regulatory T cells using diphtheria toxin-based transgenic mice. The research proposed in this application will use an innovative multidisciplinary approach that employs powerful immunology techniques rarely used to study reproductive physiology, 3D confocal microscopy, transcriptomics analysis, flow cytometry, cell sorting and bioinformatic analysis, as well as the use of mouse models to describe how specialized epithelial cells and immunocytes communicate in the epididymis. Notably, we will provide an in- depth characterization of three specific epididymal cell types: CCs and two subsets of MPs. Our proposed research is significant because it is expected to provide new insights into major immunoregulatory mechanisms by which epididymal CCs together with MPs protect spermatozoa against harmful antigens and autoimmunity. Ultimately, such knowledge will fill important gaps related to male reproductive biology by addressing crucial concepts of mucosal immunology and cell– cell interactions, all of which are critical but understudied facets of human male reproductive health.

项目摘要/摘要 男性生殖生理学最吸引人和最了解的方面之一是附子症的能力 防止针对自身抗原精子的免疫复杂发育，而启动非常有效的免疫 针对病原体的反应。该应用的长期目标是为共同开发新的战略疗法 诸如男性不育症和附睾炎等疾病，并确定男性控制的新靶标。总体 目的是阐明附睾上皮细胞（CC）如何与免疫细胞通信以有助于 “免疫特权”环境，最适合精子成熟和存储。我们最近显示了 CC在免疫激活和精子耐受性中的完全新作用。重要的是，这些细胞建立亲密 与附睾中单核吞噬细胞（MPS）的区域特异性异质子集接触。我们的中心 假设是，CC在战略上可以与免疫细胞协同工作，以调查 附睾屏障并调节炎症和免疫耐受性之间的平衡 环境。理由是基于我们的初步数据，表明CCS和MPS以稳定状态进行通信 附子症以及在感染或损伤的存在下，两种细胞类型都表达了不同的耐受性介体 允许调节精子耐受性。因此，这些细胞在附睾粘膜中起着核心作用。在 第一个具体目的，我们将确定在附睾炎期间启动免疫激活的CC - MP通信网络。 在此目的中，将使用两种小鼠的附睾炎模型：将细菌抗原注射到Cauda Lumen中，并将其注入 附睾性扭转可诱导无菌注射。在第二个特定目标中，我们将评估CC -MP 在附加症中消融精子耐受性后的串扰，通过使用白喉诱导调节T细胞耗尽 基于毒素的转基因小鼠。本应用程序中提出的研究将使用创新的多学科方法 员工的强大免疫学技术很少用于研究生殖生理学，3D共聚焦显微镜， 转录组学分析，流式细胞术，细胞分选和生物信息学分析，以及使用小鼠模型 描述专门的上皮细胞和免疫细胞如何在附睾中传达。值得注意的是，我们将提供一个 三种特定附子细胞类型的深度表征：CCS和两个MPS子集。我们提出的研究是 重要的是因为预计它将提供有关附睾的主要免疫调节机制的新见解 CC与MPS一起保护精子免受有害抗原和自身免疫性。最终，这样的知识将 通过解决粘膜免疫学和细胞的关键概念，以填补与男性生殖生物学有关的重要空白 细胞相互作用，所有这些都是至关重要的，但是人类男性复制健康方面的理解方面。