肿瘤抗原联合CD40L致敏树突状细胞诱导CTLs杀伤K562细胞的实验研究  

Antigen-loaded dendritic cells and CD40L triggers the killing effects of cytotoxic T lymphocytes on K562 cells in vitro

作　　者：林东军[1] 方志刚[1] 付咏梅[2] 李旭东[1] 黄仁魏[1] 

机构地区：[1]中山大学附属第三医院血液科,广东广州510630 [2]暨南大学医学院病理生理教研室,广东广州510632

出　　处：《中国病理生理杂志》2006年第9期1751-1755,共5页Chinese Journal of Pathophysiology

基　　金：广州市科技计划项目资助(No.2002Z3-E0012)

摘　　要：目的:以健康人外周血单核细胞为前体细胞,体外诱导为树突状细胞(DCs),负载K562细胞冻融抗原,并联合CD40L诱导产生特异性细胞毒性T淋巴细胞(CTLs)对K562细胞的杀伤作用。方法:密度梯度离心法、贴壁法分离健康人外周血单核细胞,应用rhGM-CSF、rhIL-4、rhTNF-α等细胞因子诱导扩增,培养DCs,用K562细胞冻融抗原联合rhsCD40L致敏DCs。实验分4组:K562细胞冻融抗原致敏DCs为实验组A,联合CD40L致敏DCs为实验组B,未致敏DCs为对照组A,单核细胞+异体淋巴细胞组为对照组B,观察CTLs对K562细胞的杀伤效应。结果:培养出具有典型特征的DCs,表达CD40最高达96%、CD86达97%、CD80为77%、CD1a为69%,体外能诱导强烈的同种异体混合淋巴细胞增殖反应。在效靶比为20∶1时,实验组A对K562细胞的杀伤率为71.3%,实验组B为86.9%,对照组A为37.6%,对照组B为21.1%。实验组均显示高水平杀伤率,与对照组比较差异显著(P<0.05),实验组B加CD40L杀伤率高于实验组A(P<0.05)。结论:K562细胞冻融抗原冲击致敏DCs能有效诱导T细胞特异性抗白血病作用,联合CD40L能增强其CTL的杀伤作用。AIM ： To investigate the anti - tumor effects of special cytotoxic T lymphocytes （CTLs） activated by dendritic cells （DCs） loaded with antigens and CD40L in vitro. METHODS： Peripheral blood mononuclear cells were isolated by Ficoll density gradient centrifugation from normal human heparinized blood. The adherent cells were cultured with granulocyte - macrophage colony stimulating factor （ GM - CSF）, interleukin - 4 （ IL - 4 ）, alpha tumor necrosis factor （TNF- α）, DCs were co -cultured with frozen -thawed antigen of K562 cells and CD40L, then triggered T cells into specific CTLs. RESULTS： Most suspended cells exhibited distinctive morphological features of DCs which expressed CD40 96%, CD86 97%, CD80 77%, CD1a 69%, and gained the powerful capacity to stimulate proliferation of allogenic lymphocytes. Under the effector： target ratio of 20： 1, CTLs derived from cultures with DCs and frozen - thawed antigen of K562 cells were showed 71.3% cytotoxicities against K562 cells. CTLs derived from cultures with DCs loaded with frozen - thawed antigen and CD40L were showed 86.9% cytotoxicities against K562 cells. Cytotoxicities by CTLs derived from cultures with unloaded DCs against K562 cells were 37.6% and cytotoxicities by monocytes were 21.1%. Cytotoxicities by CTLs derived from experiment groups were stronger than control groups （P 〈 0. 05 ）. CONCLUSIONS： The tumor antigen -pulsed DCs induces efficient and specific anti - tumor immunity, CTLs derived from cultures containing DCs pulsed with CIMOL show the strongest cytolytic activities on K562 cells.

关 键 词：树突细胞 冻融抗原 CIMO配体 K562细胞 T淋巴细胞 细胞毒性 

分 类 号：R363[医药卫生—病理学]