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 抗体细胞毒活性无显著变化，加入HLA-I 抗体或HLA-II 抗体对正常乳腺细胞的杀伤性均无影响。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。

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.

辽宁省科技厅社会发展攻关计划资助项目(No.2011415052- 3)