摘要
目的 探讨树突状细胞 (DC)在体外诱导针对白血病细胞的CTLs杀伤活性。方法 用IL 4和GM CSF培养正常人外周血DC ,以HL 6 0细胞裂解液致敏DC ,再与淋巴细胞共孵育 ,激活T淋巴细胞 ,用LDH释放法测定致敏DC诱导杀伤性细胞对HL 6 0细胞的杀伤活性。结果 外周血单个核细胞经体外扩增培养出可见树突状形状的DC ,流式细胞仪检测细胞表面CD1α及CD86表达阳性 ,CD14表达阴性。混合淋巴细胞反应 (MLR)观察DC刺激的异体淋巴细胞增殖明显较对照组高 ,两者比较有显著性差异 (n =15 ,P <0 .0 1)。在效靶比为 2 0∶1时 ,抗原致敏的DC诱导的杀伤性T细胞对HL 6 0细胞的杀伤率是 39.9%± 2 1.5 % ,直接以抗原刺激的T细胞的杀伤率是 2 6 .3%± 15 .6 %。两组比较差异有非常显著性 (n =16 ,P <0 .0 0 1)。效靶比为 2∶1时 ,抗原致敏的DC诱导的杀伤性T细胞对靶细胞就有明显的杀伤作用 ,随着效靶比增加 ,杀伤作用增强。实验组和对照组不同效靶比杀伤率比较 ,差异有非常显著性 (n =16 ,P <0 .0 0 1)。结论 DC在体外经HL 6 0细胞裂解液抗原致敏后 ,可诱导产生对HL 6 0细胞有特异性杀伤作用的效应细胞 ,杀伤HL 6 0细胞 ,且随效靶比的增加 。
Objective To investigate dendritic cells (DCs) pulsed with tumor antigen inducing cytotoxity of CTLS against HL 60 leukemic cells. Methods DCs were cultured in vitro with IL 4 and GM CSF from peripheral blood mononuclear cell and pulsed with HL 60 cell lysate. The phenotypes of DCs were analyzed by using flow cytometry with examining CD1α and CD86. Mixed leukocyte responses were performed by 3H intake. T cells were stimulated by DCs pulsed with antigen and T cell cytotoxicity was measured by LDH assay.Results DCs were induced from peripheral blood mononuclear cell. CD1α and CD86 is positive of the induced DCs,while CD14 is negative. Mixed leukocyte responses induced by DC was more stronger than induced by lymphocyte( n =15, P < 0.01 ). The cytotoxicity rate of 39.9% ± 21.5% was noted with DC stimulated T cells against HL 60 cells at an effector: target ratio of 20∶1 when DCs pulsed with HL 60 cell lysate, while it was 26.3% ± 15.6% for T cells pulsed with HL 60 antigen. The cytotoxity activity was stronger when the effector: target ratio got greater.Conclusion After DCs were pulsed with HL 60 lysate antigen,it can stimulate T cells proliferation and induced cytotoxity activity to kill HL 60 cell. As the effector: target ratio got greater the cytotoxity activity got stronger. It is implied that DCs pulsed with leukemia antigen can be used as immunotherapy for leukemia.
出处
《肿瘤防治研究》
CAS
CSCD
2004年第6期351-353,共3页
Cancer Research on Prevention and Treatment
基金
广东省卫生厅医学科学研究基金资助项目
