特异诱导脐血CD8^+抗白血病细胞毒淋巴细胞的研究  

Specific induction of anti-leukemia effects by umbilical cord cell-derived CD8^+T cytotoxic lymphocytes

作　　者：刘芯[1] 谭获[1] 王春燕[1] 黄振倩[1] 张还珠[1] 

机构地区：[1]广州医学院第一附属医院肿瘤血液中心,510230

出　　处：《中华血液学杂志》2006年第7期452-455,共4页Chinese Journal of Hematology

基　　金：广东省自然科学基金(020721)

摘　　要：目的研究体外诱导脐m CD8’细胞毒性T淋巴细胞(CTL)特异性杀伤门m病细胞的作用,探讨脐m淋巴细胞用于特异性免疫治疗的可行性i方法联合细胞因子体外诱导】O份脐m单个核细胞(MNC)分化为树突细胞(DC),同时负载U937冻融抗原;成熟DC刺激同源的脐m T淋巴细胞成为CTL,经MACS磁珠分选出CD8’CTL倒置显做镜、扫描电予硅微镜及流式细胞术等厅法检测DC细胞特性..MTT法测定细胞杀伤活性。结果 10份脐fli【标本均可培养出形态典型、功能成熟的DC?CD8’CTL、C1)8一CTL和淋巴细胞(TL)组对U937细胞不蚓效靶比的杀伤率以CD8’CTI.组景高;CD8’CTl。存40:l效靶比时对靶细胞U937细胞的杀伤率高于对K562细胞的杀伤率[分别为(66.36±12.43)％和(41.97±14.24)％,(P<0.05)1.而CD8 CTL在40:l效靶比时对U937细胞干“K562细胞的杀伤率差异无统汁学意义f分别为(34.47±8.19)％和(22.45±4.()O)％(尸>O.05)]结论用负载U937细胞抗原的成熟脐InL DC,诱导dj脐山l_淋巴细胞特异性的CD8’CTL;CD8’CTL对U937细胞的杀伤活性强于CD8一CTI.;CD8’CTI,对U937细胞的杀伤活性具有特异性。Objective To explore the specific anti-leukemia immune response of CD8^＋ cytotoxic T lymphocyte（CTL） derived from cord blood （CB） ex vivo and evaluate the feasibilities and values of the CTL for specific immunotherapy. Methods Dendritic cells （ DC ） were induced from mononuclear cells （ MNC ） by combination cytokines in 10 CB samples. Loading 13937 cell lysate antigen on the mature DC, they could stimulate the lymphocytes of the same origin to generate CTL. MidiMACS was used to isolate CD8 ^＋ CTL. Analysis of DC was performed by inverted microscopy, scanning electron microscopy and flow cytometry. Methyl thiazolyl tetrazolium（MTI＇） assay was used to evaluate the cytotoxicity of the CTL. Results Cocuhured with GM-CSF,IL-4,TNF-α and PGE2, CB-MNC could be induced into functional DC with typical morphology. The mean cytotoxicity of CD8^＋ CTL to U937 cells was significant stronger than that of CD8^- CTL and TL at the same E： T ratios. The mean cytotoxicity rate of CD8^＋ CTL to U937 cells was higher than that to K562 cells [（66.36±12.43）% vs （41.97±14.24）%] at E： T ratio of 40： 1（P〈0.05）. The cytotoxicity of CD^- CTL to K562 cells showed no difference from that to U937 cells （ P 〉 0.05 ）. Conclusion Mature CB-DC loading U937 cell antigens could induce CB-T lymphocytes to gengerate leukemia-specific CD8 ^＋ CTL. The cytotoxicity of the CD8^＋ CTL is specific against U937 cells and is more potent than that of CD8^- CTL.

关 键 词：T淋巴细胞 细胞毒性 免疫疗法 白血病 胎血 树突细胞 

分 类 号：R733.7[医药卫生—肿瘤]