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 |
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引用本文: | 李帆,姬静,史鹏燕,董文娟,刘勇.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[J].实用癌症杂志,2017(1):12-15. |
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作者姓名: | 李帆 姬静 史鹏燕 董文娟 刘勇 |
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作者单位: | 1. 518000,深圳益世康宁生物科技有限公司;2. 518000 深圳益世康宁生物科技有限公司;518000 深圳市第二人民医院;3. 518000,深圳北京大学香港科技大学医学中心 |
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基金项目: | 深圳市战略新兴产业发展专项资金(JSGG20141118110447437) |
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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6×1010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.
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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 |
Research on MUC1 Antigen-specific Cytotoxic T Lymphocytes for Breast Cancer |
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Abstract: | 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 ×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 's effectiveness in killing breast cancer cell provides a new option in breast cancer treatment. |
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Keywords: | Dendritic cells Cytotoxic T lymphocytes MUC1 Adeno-associated virus Breast cancer |
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