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MGBA负载脐带血DC诱导特异性CTL对乳腺癌细胞的杀伤作用[J].胡园园,刘佩佩,姜又红.中国肿瘤生物治疗杂志,2017,24(12):1381~1385.
MGBA负载脐带血DC诱导特异性CTL对乳腺癌细胞的杀伤作用    点此下载全文    点此浏览HTML全文
胡园园  刘佩佩  姜又红
中国医科大学附属第一医院肿瘤研究所,辽宁 沈阳 110001,中国医科大学附属第一医院肿瘤研究所,辽宁 沈阳 110001,中国医科大学附属第一医院肿瘤研究所,辽宁 沈阳 110001
基金项目:辽宁省科技厅社会发展攻关计划资助项目(No.2011415052- 3)
DOI:10.3872/j.issn.1007-385X.2017.12.007
摘要:
      目的:观察乳腺珠蛋白(mammaglobin A, MGBA)负载脐带血来源树突状细胞(dendritic cell, DC)诱导产生的细胞毒性T细胞(cytotoxic T lymphocyte, CTL)对乳腺癌细胞的体外杀伤效果。方法:采集健康剖宫产女性志愿者的脐带血,分离脐带血单个核细胞并诱导DC生成,用MGBA致敏DC与自体淋巴细胞共培养诱导CTL。采用流式细胞术测定DC的表型(CD83、CD86、HLADR)变化,ELISA法测定IL-10、IL-12分泌水平,CCK-8法测定CTL对乳腺癌细胞的杀伤活性。结果:成功培养出形态典型、功能成熟的DC,MGBA负载DC诱导的MGBA特异性CTL对乳腺癌细胞MDA-MB-415 产生显著的杀伤效果(P<0.05);加入HLA-I 抗体可显著减弱杀伤效果,加入HLA-II 抗体细胞毒活性显著变化,加入HLA-I 抗体或HLA-II 抗体对正常乳腺细胞的杀伤性均影响。结论:MGBA 能明显加强脐带血DC 诱导的CTL 对乳腺癌细胞的杀伤活性,该杀伤活性具有MHC限制性。
关键词:脐带血  树突状细胞  乳腺珠蛋白  乳腺癌细胞  细胞毒性T细胞
MGBA carrying umbilical cord blood-derived dendritic cells induces specific cytotoxic T lymphocytes against breast cancer cells    Download Fulltext
HU Yuanyuan  LIU Peipei  JIANG Youhong
Cancer Research Institute, The First Hospital of China Medical University, Shenyang 110001, Liaoning, China,Cancer Research Institute, The First Hospital of China Medical University, Shenyang 110001, Liaoning, China and Cancer Research Institute, The First Hospital of China Medical University, Shenyang 110001, Liaoning, China
Fund Project:Project supported by the Science and Technology Project of Liaoning Province (No.2011415052-3)
Abstract:
      Objective: To investigate the in vitro cytotoxicity of cytotoxic T lymphocytes (CTLs) induced from umbilical cord blood derived dendritic cells (DCs) that sensitized by mammaglobin A (MGBA) (MGBAp-DCs) against breast cancer. Methods: Umbilical cord blood (UCB) samples were obtained from healthy pregnant women during cesarean delivery and used to isolate UCB mononuclear cells to induce DC. MGBAp-DCs were co-cultured with autologous lymphocytes to induce CTL. The DC phenotypes (CD83, CD86, HLA-DR) were determined by using flow cytometry; and IL-10, IL-12 secretion levels was determined by ELISA; the killing effect of CTL against breast cancer cells was detected by CCK-8. Results: Mature DCs were successfully isolated with typical morphology and functionality characteristics. Specific CTL induced by MGBAp- DCs had a significant cytotoxic effect against breast cancer MDA-MB-415 cells (P<0.05). Cytotoxic effect was significantly weakened after adding anti-HLA-I antibody, but not with adding anti-HLA-II antibody. However, the the cytotoxicity of specific CTL on normal breast cell line MCF-10 was not influenced by the presence of anti-HLA-I or anti-HLA-II antibodies or not. Conclusion:MGBAp-DCs can effectively enhance the cytotocxity of CTL against breast cancer cells in vitro.
Keywords:umbilical cord blood  dendritic cell (DC)  mammaglobin A (MGBA)  breast cancer cell  cytotoxic T lymphocyte (CTL)
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