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  

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

作　　者：刘文佳[1] 朱浩[1] 吕瑛[1] 李振磊[1] 张松[1] 裴庆山[1] 邹晓平[1] 

机构地区：[1]南京大学医学院附属鼓楼医院消化科,南京210008

出　　处：《临床肝胆病杂志》2012年第8期592-597,共6页Journal of Clinical Hepatology

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<0.01)。结论 BCG活化的胰腺癌DC疫苗的生物活性明显增加,并可通过TRAIL这一途径直接杀伤肿瘤细胞;且经BCG活化的DC疫苗瘤内注射免疫治疗抗肿瘤作用更强。Objective To study the direct anti - tumor effects of the Mycobacterium boris 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, Pane02, 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 Pane02 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 pheno- typic increase in CD86 marker expression. Surface expression of TRAIL was also increased on BCG - activated DCs. Activated DCs sup- pressed the growth of Panc02 tumor ceils in culture, and this response was partially alleviated by anti - TRAIL antibody. In vivo, BCG - ac- tivated 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.

关 键 词：树突细胞 卡介苗 胰腺肿瘤 TNF相关凋亡诱导配体 

分 类 号：R735.9[医药卫生—肿瘤]