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Immunological Function of HLA-G Exepressed on trophoblasts.

滋养细胞上表达的 HLA-G 的免疫功能。

基本信息

批准号：
07671815
负责人：
ISHITANI Akiko
金额：
$ 1.47万
依托单位：
Nara Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671815/
关键词：
HLA-G placenta trophoblast monoclonal antibody immunohistochemistry immunosupressor reproductive immunology

项目摘要

To understand the basic relationship of HLA class I to maternal-placental immune tolerance, we have attempted to dissect the expression of soluble (S) HLA-G from menbrance bound (M) HLA-G and to investigate the expression of other nonclassical class I,specifically HLA-E.Towards this goal, we have developed a set of HLA-G specific monoclonal antibodies which allow the distinction of the soluble and membrane G1 proteins, and have used these and specific anti-HLA-E reagents in immunohisto-chemical staining to localize these three proteins in placental tissue. Further, we used HLA-G specific antibodies to isolate S・HLA-G and M・HLA-G directly from placental tissue in order to examine bound peptides.1. Expression of HLA-G and HLA-EImmunohistochemical staining of placenta demonstrated that S・HLA-G was found expressed in syncyciotrophoblasts, cytotrophoblasts, and extravillous cytotrophoblasts and was also apparent in maternal blood at intervillous space. Consistent with previous reports, M・HLA-G was found expressed only in extravillous cytotrophoblasts. HLA-E was expressed weakly in synciciotrophoblasts and cytotrophoblasts and comparatively more strongly in extravillous cytotrophoblasts, essentially in parallel with total HLA-G.2. Antigen presenting ability of HLA-GTo investigate the ability, TAP dependency of HLA-G expression, and sequence of HLA-G bound peptide were examined. When HLA-G was isolated from transfected cells, a relatively large number of bound peptides derived from intra cellular proteins were identified, but the number of bound peptides was considerably smaller and the individual abundance considerably higher, with a single peptide species accounting for 15% of the total peptide bound to HLA-G from the placenta.

为了理解HLA I类与母体-胎盘免疫耐受的基本关系，我们试图将可溶性（S）HLA-G的表达与膜结合（M）HLA-G分开，并研究其他非经典I类，特别是HLA-E的表达。为此，我们开发了一组HLA-G特异性单克隆抗体，其允许区分可溶性和膜G1蛋白，并在免疫组织化学染色中使用这些和特异性抗HLA-E试剂来定位胎盘组织中的这三种蛋白质。此外，我们使用HLA-G特异性抗体直接从胎盘组织中分离S·HLA-G和M·HLA-G，以检查结合肽。HLA-G和HLA-E的表达胎盘组织免疫组化染色显示，S·HLA-G在合体滋养细胞、细胞滋养细胞和绒毛外细胞滋养细胞中均有表达，在绒毛间隙母血中也有表达。与以前的报道一致，发现M·HLA-G仅在绒毛外细胞滋养细胞中表达。HLA-E在合体滋养细胞和细胞滋养细胞中表达较弱，在绒毛外细胞滋养细胞中表达相对较强，基本上与总HLA-G平行。HLA-G的抗原提呈能力为了研究这种能力，检测了HLA-G表达的TAP依赖性和HLA-G结合肽的序列。当从转染细胞中分离HLA-G时，鉴定出相对大量的来自细胞内蛋白的结合肽，但是结合肽的数量相当少，并且个体丰度相当高，单一肽种类占与来自胎盘的HLA-G结合的总肽的15%。