Analysis of the non-self recognition mechanism of ascidian hemocytes

海鞘血细胞非自我识别机制分析

基本信息

批准号：
11680687
负责人：
AZUMI Kaoru
金额：
$ 2.3万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

1) Analysis of gene expression and signal transduction pathway of ascidian hemocytes : Hemocyte aggregation is an early inflammatory response upon injury in the ascidian Halocynthia roretzi. We have already reported that the monoclonal antibody A74 strongly inhibits hemocyte aggregation in H.roretzi. The A74 antigen protein is a novel membrane glycoprotein, and it has two immunoreceptor tyrosine-based activation motifs (ITAMs) and several motifs that have been proposed to play a role in signal transduction. To identify the proteins that function downstream of ITAMs in H.roretzi hemocytes, we carried out a differential display analysis to evaluate mRNAs differentially expressed before and after hemocyte aggregation. Four amplicons (16A, 18A, 20A and 20G-1), the expression levels of which increased after induction of hemocyte aggregation, were isolated. Their expressions were found to be induced by treatment with A74 antibody, but not by control mouse IgG, and were strongly inhibited by … More treatment with BAPTA-AM, Wortmannin and cyclosporin A.From the results of cDNA cloning, it was revealed that 18A-1 clone encoded a glutathione S-transferase omega (GSTO) protein, and 20G-1 clone encoded a novel protein that had little similarity to other proteins. We also found that GSTO gene expression was specifically induced in mouse T-cell hybridoma by treatment with anti-TCR antibody.2) Analyses of opsonic factors in H.roretzi plasma and complement 3 receptor of H.roreti hemocytes : H.roretzi plasma contains two opsonic factors, galactose-specific lectin (Gal-lectin) and complement 3. Using their antibodies and a fluorescence-activated-cell sorter, we found that Gal-lectin bound only to SRBC, while C3 bound only to yeast cells. Furthermore, we found that H.roretzi hemocytes had an integrin α subunit protein and that its antibody inhibited C3-dependent phagocytosis of H.roretzi hemocytes. These results suggest that H.roretzi hemocytes have an integrin-type receptor that functions as a C3 receptor. Less

1）分析海囊血细胞的基因表达和信号转导途径：血细胞聚集是腹膜halocynthia roretzi损伤后的早期炎症反应。我们已经报道说，单克隆抗体A74强烈抑制H.Roretzi中的血细胞聚集。 A74抗原蛋白是一种新型的膜糖蛋白，它具有两个免疫感受器酪氨酸基于酪氨酸的活化基序（ITAMS），并且已经提出了一些在信号传递中发挥作用的基序。为了鉴定H.Roretzi血细胞中ITAMS下游功能的蛋白质，我们进行了差异显示分析，以评估血细胞聚集前后的mRNA不同表达。分离了四个放大器（16A，18A，20A和20G-1），分离出血细胞聚集后的表达水平升高。发现它们的表达是通过用A74抗体治疗而不是通过对照小鼠IgG诱导的，并且通过…对Bapta-Am，Wortmannin和Cyclosporin进行了更大的治疗。与其他蛋白质几乎没有相似之处。我们还发现，通过用抗TCR抗体处理的小鼠T细胞杂交瘤特异性诱导了GSTO基因表达。2）对H.Roretzi血浆中Opsonic因子的分析和H.Roretzi plasma的补体3受体包含两个Opsonic因子，包括两个Opsonic因子，使用了半乳糖 - 旋律 - 旋转 - 静脉 - 固定式（galecor-colore-colore-colore-life-colore-colore-clote-intectiv and fluce-intectiv and ins of他们的ONSICT）。分类器，我们发现GAL型蛋白仅与SRBC结合，而C3仅与酵母细胞结合。此外，我们发现H.Roretzi血细胞具有一个整合蛋白型受体，可作为C3受体。较少的