经卡介苗活化的树突状细胞疫苗治疗胰腺癌的基础及临床运用展望——暨向2011年诺贝尔医学奖得主Ralph Steinman教授致敬

刘文佳; 朱浩; 吕瑛; 李振磊; 张松; 裴庆山; 邹晓平

经卡介苗活化的树突状细胞疫苗治疗胰腺癌的基础及临床运用展望——暨向2011年诺贝尔医学奖得主Ralph Steinman教授致敬

南京大学医学院附属鼓楼医院消化科

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中图分类号: R735.9
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- 出版日期: 2012-08-20

A fight for life Honoring Ralph Steinman′s memory through a preliminary study of in vitro and in vivo anti-tumor effects of BCG-activated dendritic cells

摘要

摘要: 目的探讨经卡介苗（BCG）活化的树突状细胞（DC）疫苗体外直接抗胰腺癌细胞Panc02的机制,以及观察经不同部位注射DC疫苗抗小鼠胰腺癌移植瘤的效应。方法从C57BL/6小鼠骨髓中诱导培养DC,并予以BCG促成熟,用流式细胞技术检测DC成熟度,CCK-8细胞计数检测DC直接抑制Panc02增殖情况,流式细胞仪检测BCG促成熟DC表面凋亡诱导配体（TRAIL）的表达量。C57BL/6小鼠皮下接种Panc02胰腺癌细胞制成荷瘤小鼠,第7 d予以DC疫苗注射免疫治疗（DC疫苗为经Panc02细胞冻融抗原致敏后BCG促成熟的DC）,分为3组:瘤内注射生理盐水组、皮下注射DC疫苗组和瘤内注射DC疫苗组,1周后再治疗1次,第2次治疗后15 d处死小鼠,观察不同治疗抗肿瘤的效果。结果 BCG活化后DC疫苗成熟度明显增加,其表型CD86表达增加;经BCG活化成熟的DC能直接抑制Panc02细胞生长,这一作用可被TRAIL抗体部分抑制,且流式细胞术检测发现成熟DC表面分子TRAIL明显高于未经BCG促成熟组未成熟DC。肿瘤体重:瘤内注射DC疫苗组<皮下注射DC疫苗组<瘤内注射生理盐水组（P&...
- 树突细胞 /
- 卡介苗 /
- 胰腺肿瘤 /
- TNF相关凋亡诱导配体
Abstract: Objective To study the direct anti-tumor effects of the Mycobacterium bovis bacillus Calmette-Guerin (BCG) -activated dendritic cell (DC) vaccine in vitro and in vivo. Methods DCs were harvested from bone marrow of C57BL/6 mice and cultured with BCG to stimulate maturation.Activated DCs were evaluated by flow cytometry (FCM) .Activated DCs were co-cultured with the pancreatic tumor cell line, Panc02, and cytotoxic response was measured by the CCK-8 kit.The frequency of activated DCs expressing tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) was measured by FCM, and TRAIL mRNA levels were detected by real-time PCR.C57BL/6 mice were implanted with Panc02 tumor cells by subcutaneous inoculation, and on the seventh day were randomly divided into three groups: untreated control, injected with saline;DC vaccine-treated by intratumoral injection;DC vaccine-treated by subcutaneous injection.For all mice, injections were delivered twice with a one-week interval.On day 15 after the second injection, all the mice were sacrificed to evaluate tumor weights. Results In vitro, the BCG-activated DCs showed obvious typical dendritic structures and the phenotypic increase in CD86 marker expression.Surface expression of TRAIL was also increased on BCG-activated DCs.Activated DCs suppressed the growth of Panc02 tumor cells in culture, and this response was partially alleviated by anti-TRAIL antibody.In vivo, BCG-activated DCs decreased the Panc02-derived tumor weights (vs untreated group) , and intratumoral injection produced lower tumor weights than subcutaneous injection. Conclusion The efficacy of DC-based vaccines for pancreatic cancer may be improved by first activating the DCs with BCG.BCG-activated DCs can directly suppress the growth of Panc02 tumor cells in vitro through TRAIL.Intratumoral injection of the BCG-activated DC vaccine has a more robust anti-tumor effect than the subcutaneous route.
- dendritic cells /
- BCG vaccine /
- pancreatic neoplasms /
- TNF-related apoptosis-inducing ligand

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