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Physiology and pathophysiology of human Interleukin 12 receptor system

人白细胞介素12受体系统的生理学和病理生理学

基本信息

批准号：
10670408
负责人：
KAWASAKI Hiroshi
金额：
$ 0.64万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670408/
关键词：
cytokine chemokine receptor hybridoma genetic engineering signal transduction

项目摘要

The T cell activation requires occupancy of T cell antigen receptors (TCR) by antigenic peptide presented by Major Histocompatibility Complex (MHC) and engagement of costimulatory molecules by appropriate ligands. We were especially interested in the mechanisms of antigen presentation by antigen presenting cells (APC). To address this issue, we started to identify human chemokine receptors expressed in professional APC, dendritic cells (DC), and to clarify the involvement of chemokine receptors in antigen presentation. Several investigators reported the expression of various chemokine receptors in DC at message level. To verify these observation at protein level, we established monoclonal antibodies to a panel of C-C and C-X-C type chemokine receptors. All antibodies have been characterized whether or not inhibit ligand-induced Ca mobilization. All of them were shown to immunoprecipitate appropriate receptor molecules from physiological blood cells. We could show expression of CCR-1, CCR-3, CCR-5, CCR-6, CCR-7 and CXCR-4 in DC by fiowcytometric analysis. Of monoclonal antibodies, those amtibodies of neutralizing character against human CCR-1 and CCR-3 could strongly inhibit ligand-induced chemotaxis of DC in trans-well migration analysis, which indicates RANTES, a ligand common to CCR-1 and CCR-3 was a major chmoattractant for DC. Inhibition of DC-expressed CCR-1 and CCR-3 resulted, unexpectedly, in inhibition of allogeneic T cell stimulation by DC. The reason for inhibition may be several fold, but the most striking phenomenon observed was that DC whose CCR-1 and CCR-3 were blocked were unable to mount heterotypic aggregation with target T cells required for mounting the contact of TCR and peptide on MHC.

T细胞活化需要由主要组织相容性复合物（MHC）呈递的抗原肽占据T细胞抗原受体（TCR），并通过适当的配体接合共刺激分子。我们特别感兴趣的是抗原呈递细胞（APC）的抗原呈递机制。为了解决这个问题，我们开始鉴定在专职APC、树突状细胞（DC）中表达的人趋化因子受体，并阐明趋化因子受体参与抗原呈递。一些研究者报道了多种趋化因子受体在DC中的表达。为了在蛋白质水平验证这些观察结果，我们建立了一组C-C和C-X-C型趋化因子受体的单克隆抗体。所有抗体均已表征是否抑制配体诱导的Ca动员。所有这些都显示出从生理血细胞中免疫沉淀适当的受体分子。流式细胞术检测DC中CCR-1、CCR-3、CCR-5、CCR-6、CCR-7和CXCR-4的表达。在单克隆抗体中，对CCR-1和CCR-3具有中和作用的抗体能强烈抑制配体诱导的DC趋化性，表明CCR-1和CCR-3的共同配体RANTES是DC的主要趋化因子。DC表达的CCR-1和CCR-3的抑制意外地导致DC对同种异体T细胞刺激的抑制。抑制的原因可能是几倍的，但观察到的最显著的现象是，CCR-1和CCR-3被阻断的DC不能与靶T细胞进行异型聚集，而靶T细胞是使TCR和肽接触MHC所必需的。