SOLUBLE ISOFORMS OF HLA-G: STRUCTURE, REGULATION AND FUNCTION

HLA-G 可溶性异构体：结构、调节和功能

• 批准号: 7699703
• 负责人: JOAN Sherar HUNT
• 金额: $ 22.05万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7699703
• 关键词: Address; Affect; Age; Alternative Splicing; Antigens; Binding; Blood; Cells; Chorion; Class; Collagen Type IV; Condition; Couples; Culture Media; Cytembena; Cytotoxic T-Lymphocytes; Data; Decidua; Depth; EGF gene; Embryo; Factor V; Fertility; Fibronectins; GTP-Binding Proteins; Gene Expression; Genes; Genetic Polymorphism; Glycoproteins; Goals; HLA Class I Genes; HLA G antigen; HLA-G5; Host Defense; Human; Immune; Immune response; Immunosuppressive Agents; In Vitro; Individual; Infection; Kidney; Knowledge; Laboratories; Laminin; Ligands; Literature; Maps; Maternal-Fetal Exchange; Membrane; Messenger RNA; Monoclonal Antibodies; Mononuclear; Mothers; Normal tissue morphology; Oxygen; Pathology; Pathway interactions; Pattern; Phagocytes; Placenta; Pre-Eclampsia; Pregnancy; Pregnancy Maintenance; Premature Labor; Process; Production; Property; Protein Isoforms; Proteins; Public Health; Publications; Purpose; Reagent; Recombinant Proteins; Recurrence; Regulation; Research; Rest; Role; Sampling; Serum; Signal Transduction; Single Nucleotide Polymorphism; Spontaneous abortion; Structure; Study Section; TNFSF10 gene; Testing; Therapeutic; Tissues; Transcript; Transforming Growth Factors; Woman; Work; abstracting; base; cytokine; deprivation; design; embryo culture; embryo/fetus; experience; expression vector; insight; migration; novel; programs; receptor; research study; success; trophoblast

ABSTRACT - PROJECT I Successful human pregnancy is believed to rely upon production of HLA-G proteins by trophoblast cells in placentas. The HLA-G gene generates multiple transcripts that encode four membrane and three soluble proteins. Two of the soluble isoforms, HLA-G5 (G5) and HLA-G6 (G6) (also known as sHLA-G1 and SHLA-G2, respectively), are not only present at the maternal-fetal interface but also circulate in maternal blood throughout pregnancy. These proteins appear to be biologically important: the scientific literature documents associations between pregnancy success and high levels of soluble HLA-G in the supernatant culture media of in vitro cultured embryos, and critical preliminary experiments in our laboratory indicate that women who suffer recurrent spontaneous abortions not only fail to increase their serum levels of HLA-G5 but may decrease their levels of HLA-G6 with pregnancy . In order to study the impact of these proteins on the mother's immune responses to her genetically different embryo/fetus, we developed eukaryotic expression vectors and generated monoclonal antibodies specific for the proteins. Studies using these unique reagents have demonstrated that G5 and G6 differ in primary and secondary structures. Expression studies have documented marked differences between the isoforms in their localization to specific subpopulations of trophoblast cells, experiments on regulation of expression have demonstrated both general and isoform-specific regulatory conditions, and studies on function have identified qualitative and quantitative differences between the two isoforms. In this application we propose studies that will expand our knowledge of these powerful, pregnancy-associated proteins. AIM 1 is designed to investigate potential ligand:receptor interactions in mononuclear phagocytes, mapping expression, and performing binding studies. The goal of AIM 2 is to establish mechanisms of regulation of differential expression of G5 and G6 proteins in subpopulations of trophoblast cells from early and late gestation placentas. In AIM 3 the purpose is to investigate how G5 and G6 function to program mononuclear phagocytes. We will systematically compare results on samples from 1st trimester with term placentas and results on normal placentas with age-matched samples from problem pregnancies, i.e., preeclampsia and preterm labor/delivery with and without infection. The relevance of the proposed research to public health rests on the fact that at least one in ten couples experiences fertility difficulties and a high proportion of pregnancies fail due to preterm labor associated or not with infection. One underlying cause may be aberrancies of synthesis or expression of G5/G6 or their LILRB1/LILRB2 receptors. We expect here to provide entirely novel information that is essential to the design of therapeutic strategies to address HLA-G-associated pathologies of pregnancy.

摘要 - 项目 I 人类成功怀孕被认为依赖于滋养层细胞产生 HLA-G 蛋白。 胎盘。 HLA-G 基因产生多个转录本，编码四种膜和三种可溶性 蛋白质。两种可溶性亚型，HLA-G5 (G5) 和 HLA-G6 (G6)（也称为 sHLA-G1 和 SHLA-G2， 分别），不仅存在于母胎界面，而且在母体血液中循环 怀孕。这些蛋白质似乎具有重要的生物学意义：科学文献记录了它们之间的关联 妊娠成功且体外培养胚胎的上清液培养基中可溶性 HLA-G 水平高，以及 我们实验室的关键初步实验表明，遭受反复自然流产的妇女不会 妊娠期间仅不能增加其血清 HLA-G5 水平，但可能会降低其 HLA-G6 水平。为了 研究这些蛋白质对母亲对其基因不同的胚胎/胎儿的免疫反应的影响， 我们开发了真核表达载体并生成了针对该蛋白质的特异性单克隆抗体。 使用这些独特试剂的研究表明，G5 和 G6 在初级和次级方面存在差异 结构。表达研究记录了异构体在定位方面的显着差异 对于滋养层细胞的特定亚群，表达调节实验已经证明 一般和亚型特异性调节条件以及功能研究已经确定了定性和 两种异构体之间的数量差异。在此应用中，我们提出的研究将扩大我们的研究范围 了解这些强大的、与怀孕相关的蛋白质。 AIM 1 旨在调查潜力 单核吞噬细胞中的配体：受体相互作用、绘制表达图谱并进行结合研究。 AIM 2 的目标是建立 G5 和 G6 蛋白差异表达的调节机制。 来自妊娠早期和晚期胎盘的滋养层细胞亚群。 AIM 3 的目的是 研究 G5 和 G6 如何发挥作用来编程单核吞噬细胞。我们会系统地比较 妊娠第一个月的足月胎盘样本结果以及年龄匹配的正常胎盘样本结果 来自问题妊娠的样本，即先兆子痫和早产/分娩（有或没有感染）。 拟议研究与公共卫生的相关性取决于这样一个事实：至少十分之一的夫妇 经历生育困难，并且高比例的怀孕失败是由于相关或无关的早产 与感染。一个根本原因可能是 G5/G6 或其合成或表达的异常。 LILRB1/LILRB2 受体。我们希望在这里提供对设计至关重要的全新信息 解决 HLA-G 相关妊娠病理的治疗策略。