Impact of the menstrual cycle on granulysin-mediated immunity in the human cervicovaginal tract

月经周期对人宫颈阴道颗粒溶素介导的免疫的影响

• 批准号: 10450305
• 负责人: Florian Hladik
• 金额: $ 30.89万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-03 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10450305
• 关键词: AlamarBlue; Bacteria; Biological Assay; Biological Response Modifiers; Biopsy; Blocking Antibodies; CD4 Positive T Lymphocytes; Cell Count; Cell Survival; Cells; Cervical; Chemicals; Chlamydia Infections; Chlamydia trachomatis; Clinical Research; Clinical Trials Design; Columnar Epithelium; Contraceptive Agents; Cytolysis; Detection; Effector Cell; Endocervix; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Female; Flow Cytometry; Granzyme; Growth Factor; Hemorrhage; Home; Homologous Gene; Hormonal; Hormones; Human; Immune; Immune response; Immunity; In Vitro; Incidence; Infection; Infection Control; Infertility; Inflammation; Interferon Type II; Knowledge; Lead; Luteal Phase; Luteinizing Hormone; Lymphocyte; Measures; Mediating; Membrane; Menstrual cycle; Menstruation; Meta-Analysis; Microbe; Modeling; Mus; Nanotubes; Natural Killer Cells; Ovulation; Paper; Parasites; Participant; Patients; Pelvic Inflammatory Disease; Periodicity; Phase; Phenotype; Play; Predisposition; Prevalence; Process; Progesterone; Protein Isoforms; Proteins; Public Health; Reproductive Health; Resistance development; Role; Seminal; Serum; Sexually Transmitted Diseases; Source; Suggestion; T-Lymphocyte; Testing; Therapeutic; TimeLine; Transgenic Mice; Trypanosoma cruzi; Vagina; Visit; Wild Type Mouse; Woman; Women' s Health; antimicrobial; burden of illness; cervicovaginal; chemokine; cohort; experimental study; granulysin; improved; inhibitor; microbial; novel; pathogen; pathogenic bacteria; pathogenic microbe; perforin; proliferative phase Menstrual cycle; public health relevance; reproductive tract; translational potential; urinary

PROJECT SUMMARY/ABSTRACT Granulysin (GNLY) is an understudied human protein necessary for cell-mediated killing of microbes, including intracellular bacteria. We observed that GNLY is abundant in cervicovaginal tract (CVT) secretions during the follicular phase of the menstrual cycle, but nearly absent from the luteal phase. GNLY may play a major role in CVT immunity by killing intracellular pathogens prior to infected cell lysis (thus limiting inflammation) or even without damaging the infected cell. Thus, the change in GNLY over the menstrual cycle may have important consequences for women’s health, particularly susceptibility to sexually transmitted infections (STIs). The bacterial pathogen Chlamydia trachomatis (Ct) has a large disease burden, with more than 125 million annual infections worldwide and about 10% of untreated infections progressing to pelvic inflammatory disease, a major cause of infertility. Because Ct is an intracellular bacterial pathogen, we hypothesize that GNLY- expressing immune cells contribute to control of Ct in humans. There is suggestive evidence that Ct- specific T cells may express GNLY, but GNLY has not been studied in the context of Ct. The lack of studies may be in part due to complete control of infection by IFN-γ-producing CD4 T cells in mice. However, mice lack a GNLY homologue, making inference from mouse studies about the role of GNLY in human Ct infection impossible. In fact, human GNLY-transgenic mice control infection with other bacteria better than wild-type mice that lack GNLY. Additionally, Ct infection is more commonly detected late in the menstrual cycle, when GNLY disappears from CVT secretions. In Aim 1, we will determine how GNLY shifts so dramatically across the menstrual cycle. We will follow women from the follicular to the luteal phase. We will assess GNLY expression in cells from endocervical cytobrushes and cervical and vaginal biopsies at one visit per phase, with detailed cell phenotyping. We will assess GNLY levels in daily, self-collected secretions. In Aim 2, we will investigate GNLY’s contributions to anti-Ct immunity and increased control of CT in the follicular phase. We will perform in vitro experiments using Ct alone and Ct-infected cells; purified GNLY, granzymes, and perforin; and GNLY-expressing cells. These studies will contribute to our understanding of women’s reproductive health and STI susceptibility by elucidating mechanisms of CVT immunity during the menstrual cycle. This knowledge is important for designing clinical trials (such as of hormonal, especially intravaginal, contraceptives) and Ct therapeutics.

项目摘要/摘要 颗粒素（gnly）是一种理解的人类蛋白，用于细胞介导的微生物杀死，包括 细胞内细菌。我们观察到，gnly在宫颈阴道界（CVT）分泌过程中很丰富 月经周期的卵泡相，但在黄体期几乎不存在。 gnly可能在 CVT免疫通过在感染细胞裂解之前杀死细胞内病原体（从而限制炎症）甚至 而不会损害受感染的细胞。那就是月经周期中gnly的变化可能具有重要的 对妇女健康的后果，尤其是对性传播感染的敏感性（STIS）。 细菌病原体沙丘瘤（CT）患有大疾病伯嫩，超过1.25亿 全球年度感染，约有10％的未经治疗感染发展为骨盆炎症性疾病， 不育的主要原因。由于CT是一种细胞内细菌病原体，我们假设 表达免疫电池有助于控制人类CT。有暗示的证据表明CT- 特定的T细胞可能会表达出来，但在CT的背景下尚未研究。缺乏研究 可能部分是由于小鼠中产生IFN-γ的CD4 T细胞完全控制了感染。但是，老鼠 缺乏gnly同源物，从小鼠的研究中推断了关于gnly在人CT中的作用的研究 感染不可能。实际上，人类gnly-转基因小鼠用其他细菌控制感染比 野生型小鼠缺乏gnly。另外，在月经后期更常见CT感染。 周期，当Gnly从CVT分泌物中消失时。 在AIM 1中，我们将确定gnly如何在月经周期中如此巨大的转移。我们将跟随 从卵泡到黄体期的女性。我们将评估肠内细胞中的gnly表达 每个阶段一次访问，详细的细胞表型，细胞刺激，宫颈和阴道活检。我们将 评估每日自我收集的分泌物中的gnly水平。 在AIM 2中，我们将调查GNLY对抗CT免疫的贡献，并增加对CT的控制 卵泡期。我们将单独使用CT和CT感染的细胞进行体外实验。纯化的， 颗粒酶和穿孔蛋白；和表达gnly的细胞。 这些研究将有助于我们理解妇女的生殖健康和STI敏感性 月经周期中CVT免疫的阐明机制。这些知识对 设计临床试验（例如horsemonal，尤其是静脉内，避孕药）和CT治疗。