WT1致敏树突状细胞激活CTLs对HL60细胞作用的研究  

Research of specific cytotoxic T lymphocytes response on HL60 cells induced by antigen-loaded dendritic cells in vitro

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

机构地区：[1]中山大学附属第三医院血液科,广东广州510630

出　　处：《中华肿瘤防治杂志》2006年第19期1468-1471,共4页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的研究分别负载WT1多肽抗原及HL60细胞冻融抗原的树突状细胞(DCs)产生的特异性细胞毒性T淋巴细胞(CTLs)对HL60细胞的杀伤效应。方法联合应用rhGM-CSF、rhIL-4、rhTNF-α及rh-sCD40L等细胞因子,诱导扩增自健康人外周血获取的单核细胞,培养出DCs,分别用WT1多肽抗原及冻融抗原冲击致敏,激活淋巴细胞。实验分4组WT1多肽致敏DCs为实验组A,HL60细胞冻融抗原致敏DCs为实验组B,未致敏DCs组为对照组A,单核细胞组为对照组B,LDH释放法检测CTLs对HL60细胞的杀伤作用。结果培养出具有典型特征的DCs,表达CD40达96%,CD80达77%,CD1α达69%,CD86达97%,体外能诱导强烈的同种异体混合淋巴结胞增殖反应。在效靶比为20∶1时,WT1多肽致敏DCs组时HL60细胞杀伤率为89.1%,冻融抗原负载DCs组为90.6%,未致敏DCs组为32.8%,单核细胞组为26.1%。以下多肽或冻融抗原负载DCs与对照组相比显示更强的杀伤效应,P<0.01。结论WT1多肽抗原及HL60细胞冻融抗原冲击致敏DCs均能有效诱导T细胞抗白血病作用,为临床研制DCs疫苗提供实验依据。OBJECTIVE：To investigate the effects of dendritic cells（DCs） pulsed with WT1 peptides or frozen thawed antigen of HL60 cells on stimulating the cytotoxic T lymphocytes（CTLs） to get specific anti-tumor activity in vitro. METHODS： DCs were generated from healthy human peripheral blood monocytes in the presence of recombinant human granulocyte-macrophage colony stimulating factor, recombinant human interleukin-4, recombinant human alpha tumor necrosis factor and recombinant human sCD40L, DCs were co-cultured with WT1 peptides or frozenthawed antigen of HL60 cells, and then triggered T cells into specific CTLs. RESULTS： Most suspended cells exhibit distinctive morphological features of DCs which expressed CD40 （96%）, CD80（77%）, CDlα（69%）, CD86（97%）, and gained the powerful capacity to stimulate proliferation of allogeaic lymphcytes. Under the effector ： target ratio of 20 ： 1, CTLs derived from cul tures with DCs and WT1 peptides showed 89.1 % cytotoxicity against HL60 cells, CTLs derived from cultures with DCs and frozen-thawed antigen of HL60 cells showed 90. 6% cytotoxicity against HL60 cells, CTLs derived from cultures with unloaded DCs against HL60 cells was 32.8% and cytotoxicty of monocytes was 26.1%. Cytotoxicity of CTLs derived from culture with WT1 peptides or frozen-thawed antigen loaded DCs were stronger than in Control groups, P〈0.01. CONCLUSION： The tumor antigenpulsed DCs can induce efficient and specific anti-tumor immunity, may play a great roal in clinical therapy for leukemia.

关 键 词：树突状细胞 HL60细胞 WT1多肽 冻融抗原 细胞毒作用 

分 类 号：R733.7[医药卫生—肿瘤]