The role of semen in induction of paternal-specific tolerance during pregnancy

精液在妊娠期间诱导父亲特异性耐受中的作用

• 批准号: 10559503
• 负责人: Lucia N Vojtech
• 金额: $ 84.89万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-25 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10559503
• 关键词: Activated Lymphocyte; Address; Affect; Alloantigen; Allogenic; Antigen Presentation; Antigen-Presenting Cells; Antigens; Bar Codes; Biological Assay; Blood; Cells; Cervical; Circulation; Clonal Expansion; Clonality; Coculture Techniques; Color; Conceptions; Decidua; Dual-role transvestism; Ejaculation; Enzymes; Epidemiology; Exposure to; Failure; Fathers; Female; Female genitalia; Fetus; Frequencies; Gene Expression Profiling; Generations; Genetic Transcription; Genital; Genitalia; Human; Immune; Immune Tolerance; Immune system; Immunity; Lead; Location; Lymphocyte; Major Histocompatibility Complex; Maternal-Fetal Exchange; Metabolic; Methods; MicroRNAs; Mothers; Mucous Membrane; Mus; Oligonucleotides; Pathology; Pathway interactions; Patients; Penetration; Phenotype; Play; Population; Pre-Eclampsia; Pregnancy; Pregnancy Complications; Process; Proteins; Quantum Dots; Regulatory T-Lymphocyte; Risk; Risk Reduction; Role; Seminal fluid; Sorting; Specificity; T cell receptor repertoire sequencing; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; Techniques; Technology; Tissues; Up-Regulation; Uterus; Vagina; Vesicle; Visualization; draining lymph node; effector T cell; embryo/fetus antigen; extracellular vesicles; fetal; healthy pregnancy; hypertensive; immunoregulation; in vivo; indoleamine; innovation; lymph nodes; maternal immune system; maternal morbidity; migration; mortality; neonatal morbidity; novel; programs; receptor expression; reproductive tract; response; sex; trafficking

Human pregnancy requires maternal tolerance of the fetus. Some epidemiological evidence suggests that before conception, partner-specific tolerance begins to develop through exposure to semen, which carries paternal antigens that will also be expressed by the fetus. Regulatory T cells (Tregs) play key roles in tolerance during pregnancy, but it is unclear how these cells develop in response to paternal antigens in the female mucosa. Antigen-presenting cells (APCs) are among the first cells to be exposed to paternal antigens. They sense and respond to the local microenvironment, shift maturation status, and can induce either activated or regulatory phenotypes in T cells they encounter. Semen carries a high concentration of extracellular vesicles (EV), which we and others have shown associate with, and induce markers of tolerance in APCs. We hypothesize that these vesicles deliver paternal antigens in the form of MHC molecules, and alter mechanistic pathways to generate tolerogenic APCs, which stimulate the differentiation of Tregs specific for paternal antigens. Furthermore, we predict that in pregnancies complicated by the gestational condition preeclampsia (PE), Tregs activated by antigens present in semen will be less frequent than in healthy pregnancies. In Aim 1, we will investigate how components of semen induce tolerance in APCs from vaginal and cervical tissues. We will utilize multiple methods: our recently developed 28 color APC phenotyping panel, metabolic profiling, and transcriptional analysis to define specific pathways of tolerance induction in subsets of APCs. We will also do functional studies to investigate how exposure of APCs to semen affects the suppressive function of co-cultured T cells. In Aim 2, we will examine where semen EV distribute in the mucosa after vaginal exposure. We will employ two innovative new EV tagging technologies (quantum-dot tagging and barcoded oligonucleotide tagging) to follow the penetration into tissue and in vivo trafficking of semen EV. In Aim 3, we will determine how paternal antigen specific Tregs in the decidua and blood following delivery differ between healthy pregnancies and PE. To do this, we will isolate Tregs activated by semen antigens, and assess the clonality of the activated population by T cell receptor sequencing. We hypothesize that healthy pregnancies will have greater numbers of antigen-reactive Tregs, as well as enhanced expansion of specific clones of Tregs, indicating antigen-specificity, as compared to pregnancies complicated by preeclampsia.

人怀孕需要胎儿的孕产妇耐受性。一些流行病学的证据表明，在受孕之前，特定于伴侣特定的耐受性通过暴露于精液中开始发展，该精液带有父亲抗原，这也将由胎儿表达。调节性T细胞（Tregs）在怀孕期间在耐受性中起关键作用，但目前尚不清楚这些细胞如何响应雌性粘膜中的父亲抗原。抗原呈递细胞（APC）是最早暴露于父亲抗原的细胞之一。他们感知并应对局部微环境，转移成熟状态，并可以在遇到的T细胞中诱导激活或调节表型。精液具有高浓度的细胞外囊泡（EV），我们和其他人与APC中的耐受性相关并诱导了耐受性。我们假设这些囊泡以MHC分子的形式传递父亲的抗原，并改变了产生耐受性APC的机械途径，从而刺激对父亲抗原特异的Treg的分化。此外，我们预测，在妊娠中，胎儿症状（PE）复杂化，由精液中存在的抗原激活的Treg会比健康妊娠中的频率少。在AIM 1中，我们将研究精液的成分如何诱导阴道和宫颈组织中APC的耐受性。我们将利用多种方法：我们最近开发的28种颜色APC表型面板，代谢分析和转录分析来定义APC子集中耐受性诱导的特定途径。我们还将进行功能研究，以研究APC对精液的暴露如何影响共培养的T细胞的抑制功能。在AIM 2中，我们将检查阴道暴露后精液中EV在粘膜中分布的位置。我们将采用两种创新的新EV标记技术（量子点标记和条形码寡核苷酸标记），以遵循精液EV的组织和体内运输的渗透。在AIM 3中，我们将确定健康怀孕和PE之间分娩后的父亲抗原特异性treg和血液如何不同。为此，我们将通过精液抗原激活的Treg隔离Treg，并通过T细胞受体测序评估激活群体的克隆性。我们假设健康的妊娠将具有更多的抗原反应性Treg，以及与先兆子痫复杂的妊娠相比，Tregs的特定克隆的扩展增强，表明抗原特异性。