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.

人类怀孕需要母体对胎儿的耐受。一些流行病学证据表明，在受孕之前，伴侣特异性耐受性开始通过接触精液而形成，精液携带父本抗原，胎儿也会表达这些抗原。调节性 T 细胞 (Treg) 在妊娠期间的耐受性中发挥着关键作用，但尚不清楚这些细胞如何响应女性粘膜中的父本抗原而发育。抗原呈递细胞（APC）是最先暴露于父本抗原的细胞之一。它们感知并响应局部微环境，改变成熟状态，并且可以在遇到的 T 细胞中诱导激活或调节表型。精液携带高浓度的细胞外囊泡 (EV)，我们和其他人已经证明精液与细胞外囊泡 (EV) 相关，并诱导 APC 耐受标记。我们假设这些囊泡以 MHC 分子的形式传递父本抗原，并改变机制途径以产生耐受性 APC，从而刺激对父本抗原具有特异性的 Tregs 的分化。此外，我们预测，在妊娠期并发先兆子痫 (PE) 的情况下，被精液中存在的抗原激活的 Tregs 的频率将低于健康妊娠中的频率。在目标 1 中，我们将研究精液成分如何诱导阴道和宫颈组织 APC 的耐受性。我们将利用多种方法：我们最近开发的 28 色 APC 表型分析、代谢分析和转录分析来定义 APC 子集中耐受诱导的特定途径。我们还将进行功能研究，以研究 APC 暴露于精液如何影响共培养 T 细胞的抑制功能。在目标 2 中，我们将检查阴道暴露后精液 EV 在粘膜中的分布情况。我们将采用两种创新的 EV 标记技术（量子点标记和条形码寡核苷酸标记）来跟踪精液 EV 渗透到组织中和体内的运输。在目标 3 中，我们将确定分娩后蜕膜和血液中父系抗原特异性 Tregs 在健康妊娠和 PE 之间有何不同。为此，我们将分离被精液抗原激活的 Tregs，并通过 T 细胞受体测序评估激活群体的克隆性。我们假设，与患有先兆子痫的妊娠相比，健康妊娠将具有更多数量的抗原反应性Treg，并且Treg的特定克隆的扩增增强，这表明抗原特异性。