体外诱导的粘蛋白抗原特异性细胞毒性T淋巴细胞抗乳腺癌的研究  

Research on MUC1 Antigen-specific Cytotoxic T Lymphocytes for Breast Cancer

作　　者：李帆 姬静 史鹏燕[1] 董文娟 刘勇 

机构地区：[1]深圳益世康宁生物科技有限公司,518000 [2]深圳市第二人民医院,518000 [3]深圳北京大学香港科技大学医学中心,518000

出　　处：《实用癌症杂志》2017年第1期12-15,共4页The Practical Journal of Cancer

基　　金：深圳市战略新兴产业发展专项资金(编号:JSGG20141118110447437)

摘　　要：目的制备携带粘蛋白(MUC1)抗原基因的重组腺相关病毒(AAV/MUC1),研究其感染树突状细胞(DC)所诱导的细胞毒性T淋巴细胞(CTL)特异性杀伤肿瘤细胞的活性。方法应用分子生物学方法制备高低度的重组腺相关病毒(AAV/MUC1)。AAV/MUC1体外感染外周血单核细胞,诱导分化为DC。DC与T淋巴细胞混合,刺激产生CTL。流式细胞技术检测DC和CTL的分化和功能指标,MTS方法检测CTL的杀伤活性和特异性。结果成功制备的重组病毒(AAV/MUC1)滴度为6×10^(10)拷贝/ml。感染单核细胞率为84.27%。所获得的CTL对MUC1阳性的乳腺癌细胞株的杀伤率为(46.32±0.07)%。其杀伤作用具有MUC1抗原特异性和MHC-I类分子限制性的特征。结论以MUC1抗原为靶点的CTL可有效地杀伤乳腺癌细胞,为乳腺癌提供了一条新的治疗途径。Objective To study recombinant adeno-associated virus with human MUC-1 antigen gene （ AAV/MUC1 ） , and observe anti-tumor cell effects of MUC1 antigen-specific cytotoxic T lymphocytes （CTL）stimulated by dendritic cells（DC）in-fected by AAV/MUC1.Methods High titers of infectious AAV/MUC1 were packaged with molecular biological methods.After the monocytes were infected by AAV/MUC1,the dendritic cells（DC）were induced and generated.To generate CTL the DC were mixed with T lymphocytes.Function of the DC and CTL were detected by flow cytometry.Killing activity of the CTL against the breast cancer cells was tested by MTS.Results AAV/MUC1 was successfully generated with titers at 6 ＆#215;1010 copies/ml.The in-fection rate in monocytes was 84.27%.The killing rates of the CTL were （46.32±0.07）%.The killing activity was MUC1 anti-gen-specific and MHC class I molecule-restricted.Conclusion MUC1 antigen-specific CTL ＆#39;s effectiveness in killing breast cancer cell provides a new option in breast cancer treatment.

关 键 词：树突状细胞 细胞毒性T淋巴细胞 粘蛋白 腺相关病毒 乳腺癌 

分 类 号：R737.9[医药卫生—肿瘤]