文章摘要

K-ras 突变多肽负载的DC 细胞增强CIK 细胞对胰腺癌细胞的杀伤作用

作者: 1窦春鹏, 1李奎武, 2谭广
1 辽宁省阜新市中心医院 普外一科，辽宁 阜新 123000
2 大连医科大学附属第一医院 普外二科，辽宁 大连 116011
通讯: 谭广 Email: tanguang009@sina.com
DOI: 10.3978/.10.3978/j.issn.1005-6947.2015.03.010
基金: 国家自然科学基金资助项目, 81071173

摘要

目的：观察经K-ras（12-Val）突变多肽负载的树突状细胞（DC）与细胞因子诱导的杀伤细胞（CIK） 共培养以后对胰腺癌PANC-1 细胞的杀伤作用。 方法：取健康人外周血体外诱导分别扩增出DC 和CIK；用K-ras 突变体抗原表位肽负载DC（K-ras- DC），将单纯DC 或K-ras-DC 与CIK 共培养，获得DC-CIK 或K-ras-DC-CIK。比较CIK 与K-ras-DC-CIK 的增殖活性；分别分析DC 与K-ras-DC 以及CIK 与K-ras-DC-CIK 的免疫表型差异；检测CIK、DCCIK 、K-ras-DC-CIK 上清液中IFN-γ、IL-12 的水平； 检测K-ras-DC-CIK、DC-CIK、CIK 对PANC-1 细胞的体外杀伤力。 结果：K-ras-DC-CIK 的增殖能力明显强于单纯CIK（P<0.05）；K-ras-DC 的成熟表面分子CD1a、 CD80、CD83、HLA-DR 的表达明显高于单纯DC，而K-ras-DC-CIK 细胞群的CD3+CD8+、CD3+CD56+ 表达率明显高于单纯CIK 细胞群（均P<0.05）；上清液中IFN-γ、IL-12 的水平以及对PANC-1 细胞 的杀伤力由高到低均依次为K-ras-DC-CIK、DC-CIK、单纯CIK（均P<0.05）。 结论：K-ras 突变多肽负载后能促进DC 的成熟，负载K-ras 突变多肽后的DC 能增加CIK 的增殖及对 胰腺癌细胞的杀伤作用。
关键词： 胰腺肿瘤 树突细胞 杀伤细胞，淋巴因子激活

Killing effect of CIKs on pancreatic cancer cells enhanced by DCs loaded with K-ras mutant peptide

Authors: 1DOU Chunpeng, 1LI Kuiwu, 2TAN Guang
1 The First Department of General Surgery, Fuxin Center Hospital, Fuxin, Liaoning 123000, China
2 The Second Department of General Surgery, First Affiliated Hospital, Dalian Medical University, Dalian 116011, China

CorrespondingAuthor:TAN Guang Email: tanguang009@sina.com

Abstract

Objective: To observe the killing effect of the cytokine induced killer cells (CIKs) after co-culture with dendritic cells (DCs) harboring K-ras (12-Val) mutant peptide on pancreatic cancer PANC-1 cells. Methods: DCs and CIKs were induced and enriched from peripheral blood of healthy donors, respectively. DCs were loaded with the K-ras mutant epitope peptide (K-ras-DCs), and CIKs were co-cultured with un-loaded DCs or K-ras-DCs to obtain the DC-CIKs and K-ras-DC-CIKs, respectively. The proliferative activities between CIKs and K-ras-DC-CIKs were compared, the difference in immunophenotype between DCs and K-ras-DCs as well as between CIKs and K-ras-DC-CIKs were analyzed, the IFN-γ and IL-12 levels in the culture supernatants from CIKs, DC-CIKs and K-ras-DC-CIKs were measured, and the killing abilities of CIKs, DC-CIKs and K-ras-DCCIKs on PANC-1 cells in vitro were determined. Results: The proliferative ability of K-ras-DC-CIKs was significantly greater than that of the untreated CIKs (P<0.05); the expressions of the mature surface proteins that included CD1a, CD80, CD83 and HLA-DR in K-ras-DCs were significantly higher than those in un-loaded DCs, while the expression rates of CD3+CD56+ and CD3+CD8+ in K-ras-DC-CIK cell population were significantly higher than those in pure CIK population (all P<0.05); the levels of IFN-γ and IL-12 in the cell culture supernatant, and the killing ability on PANC-1 cells from high to low order were K-ras-DC-CIKs, DC-CIKs, and pure CIKs (all P<0.05). Conclusion: K-ras mutant peptide can promote DCs maturation, and DCs harboring K-ras mutant peptide can increase the proliferation of CIKs and killing effect on pancreatic cancer cells.
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