Analysis of the pathomechanism of the demyelinating diseases and the searach for the therapy

脱髓鞘疾病的发病机制分析及治疗方法的探索

基本信息

批准号：
07670373
负责人：
HARA Hideo
金额：
$ 1.41万
依托单位：
KYUSHU UNIVERISTY
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-07670373/
关键词：
HTLV-I rolipram ICAM1 gag HTLV-1 TCR Vβ ICAM-I 細胞内シグナル

项目摘要

Previous studies reported that production of proinflammatory cytokines, such as TNF-alpha and IFN-gamma, were increased in the serum and the CSF of HAM patients. These cytokines may play an important role in the pathogenesis of HAM/TSP.We examined the effect of PDE-IV inhibitor rolipram on the cytokines production in HTLV-I infected cell lines and PBMCs of HAM patients. PBMCs and HTLV-I infected T cell lines (HUT102, MT2) were cultured with different dose of rolipram in 10% FCS/RPMI1640. Supernatants of cultured cells were harvested after five days. Cytokines were determined by ELISA kits for TNF-alpha, INF-gamma, TGF-beta. RNA were extracted from the cultured PBMSCs and T cell lines. cDNA was synthesized and amplified with primers specific for TNF-alpha, EFN-gamma, TGF-beta by RT-PCR.Rolipram inhibited TNF-alpha production by PBMCs from all HAM patients and HUT102 in dose dependent manner. Production of IFN-gamma was less strongly suppressed in some HAM patients. Concentration of TGF-beta in culture supernatants was not influenced by rolipram. These findings suggest the possibility of rolipram as potential therapy for HAM/TSP.Intercellular adhesion molecule 1 (ICAM 1) is an inducible protein ligand which is up-regulated during inflammation and is either not consitutively expressed or expressed at low levels. The expression of ICAM1 is increased HTLV-I infected T cell lines as well as in CD4+ T cells of peripheral blood lymphocytes (PBLs) from HAM/TSP patients. After PBLs of HAM patients were cultured in the presence of stimulating anti-ICAM1 antibody, the expression of HTLV-I gag protein in PBLs was observed by the immuno-histostainning and western bloting using HTLV-I antigen specific mouse monoclonal antibody. These data suggested that the signal transduction via adhesion molecule ICAM1 could induce the transcription of HTLV-I gene and this might play an important role in the pathogenesis of HAM/TSP.

先前的研究报告说，在HAM患者的血清和CSF中增加了促炎细胞因子（例如TNF-Alpha和IFN-Gamma）的产生。这些细胞因子可能在HAM/TSP的发病机理中起重要作用。我们检查了PDE-IV抑制剂Rolipram对HTLV-I感染的HAM患者的细胞因子产生的影响。 PBMC和HTLV-I感染的T细胞系（HUT102，MT2）在10％FCS/RPMI1640中用不同剂量的Rolipram培养。五天后收获培养细胞的上清液。细胞因子通过ELISA试剂盒用于TNF-Alpha，Inf-Gamma，TGF-beta。从培养的PBMSC和T细胞系中提取RNA。 RT-PCR.Rolipram合成了cDNA并用针对TNF-Alpha，EFN-Gamma，TGF-BetA的引物进行扩增，以剂量依赖性的方式抑制了所有HAM患者和HUT102的PBMC产生TNF-Alpha。在某些HAM患者中，IFN-GAMMA的产生不太受抑制。 TGF-β在培养上清液中的浓度不受Rolipram的影响。这些发现表明，rolipram作为HAM/TSP的潜在治疗可能性。间间粘附分子1（ICAM 1）是一种可诱导的蛋白质配体，在炎症期间被上调，并且不能在低水平上表达或表达。 ICAM1的表达增加了HAM/TSP患者的外周血淋巴细胞（PBL）的HTLV-I感染T细胞系以及CD4+ T细胞中的表达。在存在刺激抗ICAM1抗体的情况下培养HAM患者PBL后，通过使用HTLV-I抗原特异性小鼠单克隆抗体，通过免疫 - 固定剂和蛋白质印迹观察到HTLV-I GAG蛋白在PBL中的表达。这些数据表明，通过粘附分子ICAM1进行信号转导可能诱导HTLV-I基因的转录，这可能在HAM/TSP的发病机理中起重要作用。