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 据信成功的人怀孕依赖于滋养细胞中HLA-G蛋白的产生 胎盘。 HLA-G基因产生了多个编码四个膜和三个可溶性的转录本 蛋白质。两个可溶的同工型HLA-G5（G5）和HLA-G6（G6）（也称为SHLA-G1和SHLA-G2， 分别存在），不仅存在于母亲界面处，而且在整个母亲的血液中循环 怀孕。这些蛋白质在生物学上似乎很重要：科学文献文件之间的关联 在体外培养的胚胎的上清液培养基中，妊娠成功和高水平的可溶性HLA-G和 我们实验室中的批判性初步实验表明，遭受反复自发堕胎的妇女 只能增加其HLA-G5的血清水平，但可能会随着怀孕而降低其HLA-G6水平。为了 研究这些蛋白质对母亲对遗传不同胚胎/胎儿的免疫反应的影响， 我们开发了真核表达载体，并产生了对蛋白质特异的单克隆抗体。 使用这些独特试剂的研究表明，G5和G6在原发性和次要方面有所不同 结构。表达研究已记录了同工型在其定位中的明显差异 对于滋养细胞细胞的特定亚群，对表达调节的实验已证明 一般和同工型特异性调节条件以及功能研究都已经确定了定性和 两种同工型之间的定量差异。在此应用中，我们提出的研究将扩大我们的 了解这些强大的，妊娠相关的蛋白质。 AIM 1旨在调查潜力 配体：单核吞噬细胞中的受体相互作用，映射表达和进行结合研究。 目标2的目的是建立在G5和G6蛋白中调节差异表达的机制 来自妊娠胎盘的滋养细胞细胞的亚群。在目标3中的目的是 研究G5和G6如何编程单核吞噬细胞。我们将系统地比较 从1个三个月的样品带有胎盘的样品，并在正常胎盘上与年龄匹配的结果 问题怀孕的样本，即先兆子痫和没有感染的早产/交付。 拟议研究与公共卫生的相关性取决于这样一个事实，即至少十分之一的夫妇 经历生育困难和高比例的怀孕因早产是否与不相关的妊娠失败 感染。一个根本原因可能是G5/G6的合成或表达的差 LILRB1/LILRB2受体。我们希望在这里提供完全新颖的信息，这对于设计至关重要 针对HLA-G相关妊娠的治疗策略。