肿瘤抗原MUC1抑制骨髓来源抑制细胞产生的机制  

Mechanism of tumor antigen MUC1 in inhibiting generation of myeloid-derived suppressor cells

作　　者：马吉春[1] 姚冬明[1] 杨静[1] 陈芹[1] 邓兆群[1] 钱炜[1] 钱军[1] 陈星星[1] 安翠[1] 闻向梅[1] 郭竑[1] 林江[1] 

机构地区：[1]江苏大学附属人民医院,江苏镇江212002

出　　处：《中国生物制品学杂志》2013年第12期1743-1747,1752,共6页Chinese Journal of Biologicals

基　　金：镇江市科技支撑-社会发展项目(SH2012033);江苏大学医学临床科技发展基金项目(JLY20120012);国家自然科学基金(81172592;81270630);镇江市科技基础设施计划项目(SS2012003)

摘　　要：目的探讨肿瘤抗原MUC1抑制骨髓来源抑制细胞(myeloid-derived suppressor cell,MDSC)产生的机制。方法分离C57BL/6小鼠股骨骨髓细胞,分别与稳定表达含22个串联重复序列的全长人MUC1 cDNA片段的B16细胞(B16-MUC1)和空质粒对照B16细胞(B16-neo)培养上清共培养,流式细胞术分析MUC1对MDSC产生及MDSC表达MUC1蛋白的影响;瑞氏姬姆萨染色观察MUC1对骨髓细胞生长形态的影响;流式细胞术检测共培养的骨髓细胞中单核细胞和巨噬细胞标志物CD14和F4/80的表达及成熟单核-巨噬细胞标志物CD11b和CD68的表达。结果正常C57BL/6小鼠骨髓细胞分别与B16-neo和B16-MUC1细胞培养上清共培养24和48 h后,BM+B16-MUC1组骨髓细胞中CD11b+Gr-1+MDSC数量较BM+B16-neo组均明显减少(P<0.05),共培养48 h后,BM+B16-MUC1组的CD11b+Gr-1+MDSC中MUC1蛋白的表达较BM+B16-neo组明显增多(P<0.01);BM+B16-MUC1组骨髓细胞中单核-巨噬细胞数量增多;与BM+B16-neo组相比,BM+B16-MUC1组CD14+、F4/80+和CD68+细胞表达数量明显增高(分别为P<0.01、P<0.01、P<0.05),CD11b+平均荧光强度明显增高(P<0.01)。结论 MUC1通过诱导骨髓细胞向单核-巨噬细胞分化,从而抑制MDSC的产生。Objective To investigate the mechanism of tumor antigen MUC1 in inhibiting the generation of myeloid-derived suppressor cells （MDSCs）. Methods Marrow cells in thighbone of C57BL/6 mice were separated and co-cultured with the culture supernatants of B16-MUC1 cells stably expressing the full-length MUC1 cDNA containing 22 tandem repeat sequence and B16-neo cells containing empty plasmid as control respectively. The production of MDSCs and expression of MUC1 were determined by flow cytometry （FCM）. The morphological change of marrow cells were observed by Wright-Giemsa staining. The expressions of mononuelear and macrophage cell makers CD14 and F4/80 as well as monocyte-macrophage markers CDllb and CD68 in co-cultured bone marrow cells were determined by FCM. Results The numbers of CD11b ＋GR-1＋ MDSCs in BM ＋ B16-MUC1 group 24 and 48 h after co-culture decreased significantly as compared with those in BM ＋ B16-neo group （P 〈 0. 05）. However, the expression level of MUC1 in BM ＋ B16-MUC1 group 48 h after co-culture increased significantly as compared with that in BM ＋ B16-neo group （P 〈 0. 01 ）, while the counts of monocytes-macrophages increased, and the counts of CD14＋,F4 / 80＋ and CD68＋ cells as well as average fluorescent intensity of CD11b increased significantly（each P 〈 0. 01 except CD68 P 〈 0. 05）. Conclusion MUC1 inhibited the generation of MDSCs by inducing the differentiation of marrow cells to monocytes-macrophages.

关 键 词：MUC1 骨髓来源抑制细胞 免疫调节 肿瘤 

分 类 号：R73-362[医药卫生—肿瘤]