TJU103体外诱导造血干细胞移植的免疫耐受  

Immunotolerance to haemopoietic stem cell transplantation induced by TJU103 in vitro

作　　者：周健[1] 涂三芳[1] 郭坤元[1] 孙明[1] 胡亮杉[1] 吴远彬[1] 卢晓珣[1] 王杨[1] 

机构地区：[1]南方医科大学珠江医院血液科,广东省广州市510282

出　　处：《中国组织工程研究与临床康复》2008年第16期3049-3052,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：广州市政府科技计划项目(200523-E0551)~~

摘　　要：目的:前期研究表明,吲哚亚甲基异烟腙(简称TJU103)可以明显降低异基因造血干细胞移植小鼠移植物抗宿主病的发生率和程度,明显延长小鼠移植后的生存期。实验拟观察TJU103诱导造血干细胞移植免疫耐受和对移植物抗白血病的影响,探索一条既能降低移植物抗宿主病又能保留移植物抗白血病的移植途径。方法:实验于2007-07/10在南方医科大学珠江医院血液科实验室完成。①实验材料:人急性髓系白血病细胞株KG1a由中国医学科学院血液学研究所宋增璇教授惠赠。10份自愿捐献的健康人外周血白细胞由广州市中心血站提供,等分为供者与受者。②实验方法:采用梯度密度沉淀法常规分离人外周血单个核细胞。以供者外周血单个核细胞作为反应细胞,受者正常外周血单个核细胞作为刺激细胞,体外建立异基因造血干细胞移植中供、受者间的单向混合淋巴细胞反应体系,应用TJU103阻断CD4+T细胞激活的抗原信号通路,设单独刺激细胞和反应细胞对照以及1640完全培养基空白对照。③实验评估:于培养第1、3、5天应用MTT检测TJU103对供者淋巴细胞的增殖活性的影响,于第5天用乳酸脱氢酶释放法分别检测10∶1,20∶1,40∶1效靶比时供者淋巴细胞对受者单个核细胞和急性髓系白血病细胞KG1a的细胞毒活性影响,同时采用流式细胞仪检测CD4+CD25+T细胞的百分比。结果:第3天和第5天实验组A值低于对照组(P<0.05)。单向混合淋巴细胞5d后实验组各效靶比供者T细胞杀伤受者外周血单个核细胞的活性明显低于对照孔(P<0.05);二者对KG1a细胞的杀伤活性差异不显著(P>0.05)。TJU103对供者CD4+CD25+T细胞无影响。结论:TJU103降低T淋巴细胞增殖反应的同时并不影响其抗白血病效应,有望用于临床异基因造血干细胞移植中移植物抗宿主病和移植物抗白血病的免疫调节。AIM： Pristine investigation demonstrated that TJU103 could markedly decrease graft versus host disease （GVHD） incidence and intensity, and obviously prolonge life span after allogenetic haemopoietic stem cell transplantation （allo-HSCT） in mice. This experiment is designed to explore the effects of TJU103 on tolerance and graft versus leukemia （GVL） in allo-HSCT in vitro in order to seek an effective access to transplantation remaining GVL and with less GVHD. METHODS： Experiments were performed at the Department of Hematology of Zhujiang Hospital of Southern Medical University from July to October 2007. （1）Human acute myelogenous leukemia lines KGla was presented by professor Song in Hematology Institute of Chinese Academy of Medical Sciences. All 10 peripheral blood samples were collected from healthy volunteers at Guangzhou Blood Center, and were divided equally into donors and recipients. （2） Peripheral blood mononuclear cells （PBMCs） were obtained by traditional precipitation method discriminated by gradient density. Donor PBMCs and recipient PBMCs were took as responsive cell and antigenic stimulus cell, respectively. One-way mixture lymphocyte culture （MLC） response system between donor and recipient was modeled in vitro. The antigen activated pathway of CD4 positive T cell was blocked by TJU103. The blank control group of 1640 complete medium and control group of alone responsive cell and antigenic stimulus cell were set. （3）Effects of TJU103 on donor T lymphocytes proliferation was estimated by MTT assay at MLC 1, 3, 5 days. Cytotoxicity of donor T lymphocytes blocked by TJUI03 against recipient T lymphocytes and KGIa cell were measured by LDH releasing assay at different effector-to-target cell ratios （E：T, 10：1, 20：1 40：1） at MLC 5 days. Meanwhile, the ratio of donor CD4^＋CD25^＋ T lymphocytes was assayed by flow cytometry （FCM）. RESULTS： The A numerical values of experiment group were lower than control group at MLC 3 and 5 days （P 〈 0.05�

关 键 词：干细胞移植 TJU103 T淋巴细胞增殖 细胞毒活性 移植物抗宿主病 移植物抗白血病 免疫耐受 

分 类 号：R394.2[医药卫生—医学遗传学]