细胞因子IL-7与IL-15联合优化胃癌EBV特异性CTL细胞培养体系  

Optimization of EBV specific CTLs cell culture system with the combination of IL-17 and IL-15 cytokines in gastric cancer

作　　者：邵洁[1] 苏舒[1] 徐秋萍[1] 邹征云[1] 魏嘉[1] 杜娟[1] 孟凡岩 钱晓萍[1] 刘宝瑞[1] Shao Jie;Su Shu;Xu Qiuping;Zou Zhengyun;Wei Jia;Du Juan;Meng Fanyan;Qian Xiaoping;Liu Baorui(The Comprehensive Cancer Center of Drum Tower Hospital,Medical School of Nanjing University&Clinical Cancer Institute of Nanjing University,Jiangsu Nanfing 210008,China)

机构地区：[1]南京大学医学院附属鼓楼医院肿瘤中心,江苏南京210008

出　　处：《现代肿瘤医学》2018年第24期3902-3908,共7页Journal of Modern Oncology

基　　金：国家自然科学基金(编号:81572601;81702811)

摘　　要：目的:探讨细胞因子白细胞介素-7(IL-7)和IL-15在胃癌EB病毒(EBV)抗原特异性细胞毒性T细胞(CTLs)培养中较IL-2的优势。方法:提取胃癌患者外周血单个核细胞贴壁培养2 h,贴壁细胞加IL-4和粒细胞-巨噬细胞集落刺激因子(GM-CSF)诱导树突状细胞(DC),成熟的DC负载EBV抗原肽与T细胞混合培养,诱导EBV-CTLs,比较在细胞因子IL-2或IL-7/15培养条件下诱导的EBV-CTLs的扩增能力、表型、免疫应答及体外杀伤能力的差别。结果:与细胞因子IL-2相比较,IL-7联合IL-15培养的EBV-CTLs细胞中含有较高比例的CD3^+T细胞[(91. 2±1. 5)%vs (80. 8±1. 6)%,P=0. 008 4]以及较高比例的CD3^+CD8^+T细胞[(66. 8±2. 3)%vs (30. 17±6. 9)%,P=0. 007 6];在维持中央记忆性T细胞扩增方面,IL-7联合IL-15扩增的EBV-CTLs细胞中含有较高比例的CD8^+CD62L^+T细胞[(26. 4±2. 3)%vs(9. 6±1. 4)%,P=0. 003 6]及CD8^+CD27^+T细胞[(38. 5±3. 7)%vs (16. 7±2. 9)%,P=0. 01];在免疫应答能力方面,接触DC负载抗原肽24 h后,IL-7联合IL-15培养的EBV-CTLs能分泌更多的γ干扰素(IFN-γ),且CD137上调明显高于IL-2组;在体外杀伤实验中,IL-7联合IL-15诱导的EBV-CTLs对表达EBV抗原的靶细胞表现出更强的杀伤作用。结论:IL-7联合IL-15在扩增胃癌抗原特异性EBV-CTLs方面具有优势,扩增的细胞中含有较高比例的CD3^+CD8^+T细胞,且能维持中央记忆性T细胞的扩增,表现出有更强的免疫应答和体外抗肿瘤效果,为其进一步临床个体化治疗EBV阳性胃癌提供客观实验依据。Objective： To investigate the advantages of cytokine IL-7 and IL-15 in the development of gastric cancer EBV antigen-specific CTLs. Methods： DCs were generated from monocytes enriched by adherence for 2 h,and cultured in medium containing human GM-CSF and IL-4. Mature DCs were pulsed by EBV antigen peptide and then incubated with T cells to induce EBV-CTLs. To compare the amplification ability,phenotype,cellular immune response ability and in vitro killing ability of EBV-CTLs induced by different cytokine culture conditions. Results： Compared with IL-2,IL-7/15 expanded EBV-CTLs populations contained more CD3^＋T cells [（ 91. 2± 1. 5） % vs（ 80. 8 ± 1. 6） %,P = 0. 008 4 ],meanwhile,IL-7/15 expanded EBV-CTLs populations contained more CD3^＋CD8^＋T cells as compared with IL-2 [（ 66. 8 ± 2. 3） % vs（ 30. 17 ± 6. 9） %,P = 0. 007 6]. Maintenance of central memory T cell expansion,we observed higher proportion of CD8^＋CD62 L^＋T cell in culture with IL-7/15[（ 26. 4 ± 2. 3） % vs（ 9. 6 ± 1. 4） %,P = 0. 003 6]. Besides,we found the IL-7 combination with IL-15 led to the greater expansion of CD8^＋CD27^＋T cell[（ 38. 5 ± 3. 7） % vs（ 16. 7 ± 2. 9） %,P = 0. 01]. We found in culture with IL-7/15 led to more IFN-γ secretion and CD137 expression. EBV-CTLs cultured under IL-7 combined with IL-15 showed a stronger lysis effect than other groups. Conclusion： In this study,IL-7 combined with IL-15 promotes the expansion of EBV-CTLs with more CD3^＋CD8^＋T cells,enhanced immune response,and superior cytotoxic effector characteristics these results provide an objective experimental basis for the further individualized treatment of EBV positive gastric cancer.

关 键 词：过继性免疫治疗 细胞毒性T淋巴细胞 EBV感染 IL-7 IL-15 

分 类 号：R73-35[医药卫生—肿瘤]