小鼠3LL肿瘤局部高表达GM-CSF增强抗肿瘤免疫的实验研究  

A study on enhanced anti-tumor immunity induced by high level expression of GM-CSF in 3LL tumor site of the mice

作　　者：林怡[1] 张镭[1] 张毅[1] 蔡玉婵[1] 熊思东[1] 储以微[1] 

机构地区：[1]复旦大学上海医学院免疫学系,教育部分子医学重点实验室,上海200032

出　　处：《中国癌症杂志》2008年第1期15-19,共5页China Oncology

基　　金：国家自然科学基金面上项目(30571713);复旦大学上海医学院基础临床医学交叉基金(2007JC07-1)

摘　　要：背景与目的:粒-单核细胞集落刺激因子(grannulocyte/monocyte-colony stimulating factor,GM-CSF)具有促进未成熟树突状细胞(dendritic cell,DC)成熟、上调其MHC分子和协同刺激分子表达以及增强DC对肿瘤抗原的提呈等作用。本文研究GM-CSF在肿瘤局部的高表达可以促进肿瘤局部DC成熟及亚群改变,从而诱导增强的抗肿瘤免疫效应。方法:以GM-CSF转染小鼠Lewis肺癌细胞株3LL获得3LL-GM,分别以3LL、3LL-vec(空载体对照)和3LL-GM细胞接种C57BL/6小鼠,分离肿瘤局部免疫细胞,流式细胞术检测DC成熟度、DC亚群比例、CD8+效应T细胞的活化及其功能。结果:3LL-GM肿瘤局部GM-CSF浓度高达441.22ng/g肿瘤组织,显著高于母本3LL组的0.53ng/g肿瘤组织(P<0.05)和空载体对照3LL-vec组的0.42ng/g肿瘤组织(P<0.05)。在3LL-GM、3LL和3LL-vec组小鼠肿瘤内浸润的CD11c+DC细胞中,I-Ab+细胞的比例分别为60.62%、19.98%和23.12%(P<0.05),CD80+细胞的比例分别为60.93%、37.43%和47.03%(P<0.05),表明肿瘤局部GM-CSF促进DC成熟;同时,三组小鼠肿瘤内浸润的CD11c+CD8α+CD4-DC亚群比例分别为60.82%、40.00%和29.27%(P<0.05),显示肿瘤局部GM-CSF促使DC分化为CD11c+CD8α+CD4-亚群。3LL-GM组小鼠肿瘤内浸润淋巴细胞(tumor in-filtrating lymphocyte,TIL)中,CD3+CD62Llow细胞比例高达20.84%,显著高于3LL组的6.34%(P<0.05)和3LL-vec组的15.18%(P<0.05);并且分泌IFN-γ的CD8+T淋巴细胞比例为2.77%,与对照组相比差异具有显著性(P<0.05)。结论:肿瘤局部高表达GM-CSF,可通过提高DC成熟度及上调CD11c+CD8α+CD4-DC亚群在肿瘤局部的比例,诱导增强的抗肿瘤免疫效应,最终导致肿瘤消退。Background and purpose： Grannulocyte/monocyte-colony stimulating factor（GM-CSF） can facilitate maturation of immature dendritic cells（DC）, upregulate their MHC molecules and co-stimulating factors, as well as enhance tumor antigen presentation by DC. Our study was to investigate whether high level expression of GM-CSF in tumor site could induce enhanced anti-tumor immune response and its mechanism. Methods： 3LL-GM was established by transduction of 3LL, Lewis lung cancer cell line, with lentivirus coded with GM-CSF gene. C57BL/6 mice were inoculated subcutaneously with 3LL, 3LL-vec and 3LL-GM, respectively. DC maturation and the percentage of DC subsets, as well as the activation and function of ]ymphocytes especially CD8^＋ effector T lymphocytes in tumor site were detected. Results： The concentration of GM-CSF in 3LL-GM tumor sites was 441.22 ng/g tumor, significantly higher than in parental 3LL tumor site （0.53 ng/g tumor, P 〈 0.05） and in vector control 3LL-vec tumor site （0.42 ng/g tumor, P 〈 0.05）. In 3LL-GM group, 3LL and 3LL-vec group, percentages of I-A^b＋ cells in tumor infiltrating CDllc^＋ DCs were 60.62%, 19.98% and 23.12% respectively （ P 〈 0.05）, and percentages of CD80^＋ cells were 60.93%, 37.43% and 47.03% respectively（ P 〈 0.05）, indicating that GM-CSF in a tumor site can facilitate maturation of DCs. Meanwhile, proportions of tumor infiltrating CD1 lc^＋ CD8α^＋ CD4- DC in 3 groups were 60.82%, 40.00% and 29.27% respectively（ P 〈0.05）, showing elevated differentiation into CD1 lc^＋ CD8α^＋ CD4^- DC subset induced by local GM-CSF. Moreover, CD3^＋ CD62L^low cells in tumor infiltrating lymphocytes （TILs） of 3LL-GM group was 20.84%, significantly higher than those of 3LL group （6.34%, P 〈0.05） and 3LL-vec group （15. 18%, P 〈 0. 05）. More importantly, percentage of IFN-secreting CD8 ＋ T lymphocytes was 2.77%, showing significant statistical difference from the control group （P 〈 0.05）, and consequently caused marked

关 键 词：粒-单核细胞集落刺激因子 树突状细胞 3LL 移植瘤 小鼠 

分 类 号：R392.11[医药卫生—免疫学]