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.

先前的研究报道，HAM 患者的血清和脑脊液中促炎细胞因子（如 TNF-α 和 IFN-γ）的产生增加。这些细胞因子可能在HAM/TSP的发病机制中发挥重要作用。我们检测了PDE-IV抑制剂咯利普兰对HTLV-I感染细胞系和HAM患者PBMC中细胞因子产生的影响。将PBMC和HTLV-I感染的T细胞系(HUT102、MT2)与不同剂量的咯利普兰在10％FCS/RPMI1640中培养。五天后收获培养细胞的上清液。通过 ELISA 试剂盒测定细胞因子的 TNF-α、INF-γ、TGF-β。从培养的 PBMSC 和 T 细胞系中提取 RNA。通过 RT-PCR 合成并使用 TNF-α、EFN-γ、TGF-β 特异性引物扩增 cDNA。咯利普兰以剂量依赖性方式抑制所有 HAM 患者和 HUT102 的 PBMC 产生 TNF-α。在一些 HAM 患者中，IFN-γ 的产生受到不太强烈的抑制。培养物上清液中TGF-β的浓度不受咯利普兰的影响。这些发现表明咯利普兰有可能作为 HAM/TSP 的潜在疗法。细胞间粘附分子 1 (ICAM 1) 是一种诱导型蛋白配体，在炎症过程中上调，并且不持续表达或表达水平较低。 ICAM1 的表达在 HTLV-1 感染的 T 细胞系以及来自 HAM/TSP 患者的外周血淋巴细胞 (PBL) 的 CD4+ T 细胞中增加。在刺激性抗ICAM1抗体存在下培养HAM患者的PBL后，使用HTLV-I抗原特异性小鼠单克隆抗体通过免疫组织染色和蛋白质印迹观察PBL中HTLV-I gag蛋白的表达。这些数据表明，通过粘附分子ICAM1进行的信号转导可以诱导HTLV-I基因的转录，这可能在HAM/TSP的发病机制中发挥重要作用。