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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

基本信息

批准号：
10616798
负责人：
Florian Hladik
金额：
$ 17.65万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-03 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10616798
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 Listeria monocytogenes 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 T-Lymphocyte Testing Therapeutic Toxoplasma gondii Transgenic Mice Trypanosoma cruzi Vagina Visit Wild Type Mouse Woman Women&apos 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 timeline translational potential urinary

项目摘要

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.

项目概要/摘要颗粒溶素 (GNLY) 是一种尚未充分研究的人类蛋白质，对于细胞介导的微生物杀伤是必需的，包括细胞内细菌。我们观察到 GNLY 在宫颈阴道道 (CVT) 分泌物中含量丰富月经周期的卵泡期，但黄体期几乎不存在。 GNLY 可能发挥重要作用 CVT 免疫通过在受感染的细胞裂解之前杀死细胞内病原体（从而限制炎症）甚至而不损坏受感染的细胞。因此，月经周期中 GNLY 的变化可能具有重要意义。对女性健康的影响，特别是对性传播感染（STI）的易感性。细菌病原体沙眼衣原体 (Ct) 具有巨大的疾病负担，超过 1.25 亿人全球每年都会发生感染，约 10% 的未经治疗的感染会进展为盆腔炎，不孕不育的一个重要原因。由于 Ct 是一种细胞内细菌病原体，我们假设 GNLY- 表达免疫细胞有助于控制人类的 Ct。有暗示性证据表明 Ct- 特定的 T 细胞可能表达 GNLY，但 GNLY 尚未在 Ct 背景下进行研究。缺乏研究部分原因可能是由于小鼠体内产生 IFN-γ 的 CD4 T 细胞完全控制了感染。然而，老鼠缺乏 GNLY 同源物，从小鼠研究中推断 GNLY 在人类 Ct 中的作用感染不可能。事实上，人类 GNLY 转基因小鼠比其他小鼠更好地控制其他细菌的感染。缺乏 GNLY 的野生型小鼠。此外，Ct 感染更常见于月经后期循环，当 GNLY 从 CVT 分泌物中消失时。在目标 1 中，我们将确定 GNLY 如何在整个月经周期中发生如此巨大的变化。我们将跟随女性从卵泡期到黄体期。我们将评估宫颈管细胞中 GNLY 的表达每阶段一次进行细胞刷以及宫颈和阴道活检，并进行详细的细胞表型分析。我们将评估每日自行收集的分泌物中的 GNLY 水平。在目标 2 中，我们将研究 GNLY 对抗 Ct 免疫和增强 CT 控制的贡献卵泡期。我们将使用单独的Ct和感染Ct的细胞进行体外实验；纯化的GNLY，颗粒酶和穿孔素；和 GNLY 表达细胞。这些研究将有助于我们了解女性生殖健康和性传播感染的易感性阐明月经周期期间 CVT 免疫机制。这些知识对于设计临床试验（例如激素，尤其是阴道内避孕药）和 Ct 疗法。