抗原冲击致敏树突状细胞诱导特异性CTL杀伤Jurkat细胞实验研究  

Antigen-loaded Dendritic Cells Trigger Killing Effects of Specific Cyto-toxic T Lymphocytes on Jurkat Cells In Vitro

作　　者：林东军[1] 方志刚[1] 李旭东[1] 刘加军[1] 陆英[1] 

机构地区：[1]中山大学附属第三医院血液科,广州510630

出　　处：《中国实验血液学杂志》2006年第4期795-799,共5页Journal of Experimental Hematology

基　　金：广州市科技计划项目;编号:2002Z3-E0012

摘　　要：本研究以健康人外周血单核细胞为前体细胞,体外诱导其为树突状细胞(DC),分别负载Jurkat细胞冻融抗原及WT1多肽抗原,产生特异性细胞毒性T淋巴细胞(CTL),以探讨其对白血病Jurkat细胞的杀伤作用。联合应用rhGM-CSF、rhIL-4、rhTNF-α及rhsCD40L等细胞因子,密度梯度离心法、贴壁法从外周血获取单核细胞并进行诱导扩增,培养出DC,分别用冻融抗原及WT1多肽抗原冲击致敏DC。实验分4组冻融抗原致敏DC组为实验组A,WT1多肽致敏DC组为实验组B,未致敏DC组为对照组A,单核细胞组为对照组B,观察CTL对Jurkat细胞的杀伤作用。结果表明培养出了具有典型特征的DC,它高表达CD40、CD80、CD1a及CD86等表面标志,能体外诱导强烈的同种异基因混合淋巴细胞增殖反应。实验组显示高水平杀伤率,与对照组比较均具有统计学意义(P<0.01),实验组A、B间比较无统计学意义。结论Jurkat细胞冻融抗原及WT1多肽抗原冲击致敏DC能有效诱导T细胞抗白血病作用,为临床研制DC疫苗提供了实验依据。This study was aimed to investigate the effects of tumor antigen-loaded dendritic cells （DC） stimulating the specific cytotoxic T lymphocytes （CTL） on Jurkat cells in vitro. Peripheral blood mononuclear cells were isolated by Ficoll density gradient centrifugation from normal human heparinized blood, the adherent monocytes were cultured with granulocyte-macrophage colony stimulating factor （ GM-CSF）, interleukin-4 （ IL-4 ）, alpha tumor necrosis factor （TNF-α） and sCD40L,DCs were co-cultured with frozen-thawed antigen of Jurkat cells or WT1 peptides, and then T cells were triggered into specific CTL. The results showed that most suspended cells exhibited distinctive morphological features of DC which expressed CD40 （96%）, CD86 （97%）, CD80 （77%）, CDIa （69%）, and gained the powerful capacity to stimulate proliferation of allogeneic lymphocytes. Under the effector ： target ratio of 20：1, CTLs derived from cultures with DC and frozen-thawed antigen of Jurkat cells showed 91.1% cytotoxicity against Jurkat cells, CTL derived from cultures with DC and WT1 peptides showed 87.5% cytotoxicity against Jurkat cells, cytotoxicity of CTL derived from cultures with unloaded DC against Jurkat cells was 42.1% and cytotoxicity of monocytes was 22.7%. Cytotoxicity of CTL derived from culture with frozen-thawed antigen or WTI peptides loaded DC was stronger than that in control groups（P 〈0.01 ）. It is concluded that the tumor antigen-pulsed DC can induce efficient and specific anti-tumor immunity, may play a great role in clinical therapy for leukemia.

关 键 词：树突状细胞 JURKAT细胞 冻融抗原 WT1多肽抗原 细胞毒作用 白血病 

分 类 号：R730.3[医药卫生—肿瘤] R392.1[医药卫生—临床医学]