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

详细信息

中图分类号: R735.9
计量
- 文章访问数: 245
- HTML全文浏览量: 11
- PDF下载量: 183
- 被引次数: 0
出版历程
- 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

HTML全文

参考文献(28)

[1]Burris HA, Moore MJ, Andersen J, et al.Improvements insurvival and clinical benefit with gemcitabine as first-linetherapy for patients with advanced pancreas cancer:a ran-domized trial[J].J Clin Oncol, 1997, 15 (6) :2403-2413.

[2]Rothenberg ML, Moore MJ, Cripps MC, et al.A phase ii tri-al of Gemcitabine in patients with 5-fu-refractory pancreascancer[J].Ann Oncol, 1996, 7 (4) :347-353.

[3]Louvet C, Labianca R, Hammel P, et al.Gemcitabine in combi-nation with oxaliplatin compared with gemcitabine alone in local-ly advanced or metastatic pancreatic cancer:Results of a gercorand giscad phase iii trial[J].J Clin Oncol, 2005, 23 (15) :3509.

[4]Dauer M, Schnurr M, Eigler.A Dendritic cell-based cancervaccination:Quo vadis?[J].Expert Rev Vaccines, 2008, 7 (7) :1041-1053.

[5] Endo H, Saito T, Kenjo, et al.Phase I trial of preoperativeIntratumoral injection of immature dendritic cells and OK-432 forresectable pancreatic cancer patients[J].J Hepatobil-iary Pancreat Sci, 2012, 19 (4) :465-475.

[6]Mellman I, Nussenzweig M.Retrospective.Ralph M.Stein-man (1943-2011) [J].Science, 2011, 334 (6055) :466.

[7]Lanzavecchia A, Sallusto F.Ralph M.Steinman 1943-2011[J].Cell, 2011, 147 (6) :1216-1217.

[8]Gravitz L.A fight for life that united a field[J].Nature, 2011, 478 (7368) :163-164.

[9]Steinman RM, Banchereau J.Taking dendritic Cells Into medi-cine[J].Nature, 2007, 449 (7161) :419-426.

[10]Kim HS, Choo YS, Koo T, et al.Enhancement of antitumorimmunity of dendritic cells pulsed with heat-treated tumorlysate in murine pancreatic cancer[J].Immunol Lett, 2006, 103 (2) :142-148.

[11]Gunzer M, Janich S, Varga G, et al.Dendritic cells and tumorimmunity[J].Semin Immunol, 2001, 13 (5) :291-302.

[12]Tjomsland V, Sandstr m P, Sp ngeus A, et al.Pancreaticadenocarcinoma exerts systemic effects on the peripheralblood myeloid and plasmacytoid dendritic cells:an indicatorof disease severity[J].BMC Cancer, 2010, 10:87.

[13]徐杰, 陆华, 吴卫江, 等.人脑胶质瘤DC疫苗的临床应用[J].江苏医药, 2012, 38 (10) :1193-1195.

[14]刘刚, 庞利群.卡介苗免疫治疗肿瘤的研究进展[J].江苏大学学报 (医学版) , 2008, 18 (12) :181-184.

[15]Kim KD, Lee HG, Kim JK, et al.Enhanced antigen-presen-ting activityand tumour necrosis factor-a-independent acti-vation of dendritic cells following treatment with Mycobacteri-um bovis bacillus Calmette–Gue'rin[J].Immunology, 1999, 97 (4) :626-633.

[16]Demangel C, Bean AG, Martin E, et al.Protection againstaerosol Mycobacterium tuberculosis infection using Myco-bacterium bovis Bacillus Calmette Guerin-infected dendriticcells[J].Eur J Immunol, 1999, 29 (6) :1972-1979.

[17] 杨德红, 刘文佳, 陈敏, 等.经卡介苗活化的负载PANC1裂解产物的DC疫苗体外抗胰腺癌作用[J].中华胰腺病杂志, 2010, 10 (4) :267-281.

[18]Roux S, Apetoh L, Chalmin F, et al.CD4+CD25+Tregscontrol the TRAIL dependent cytotoxicity of tumor-infiltra-ting DCs in rodent models of colon cancer[J].J Clin Invest, 2008, 118 (11) :3751-3761.

[19] 刘文佳, 曹俊, 邹晓平, 等.DC胰腺癌免疫治疗新进展[J].中华消化杂志, 2008, 28 (8) :575-576.

[20] 刘文佳, 邹晓平, 曹俊, 等.热休克联合OK-432处理Panc-1细胞裂解物致敏的DC体外诱导特异性抗肿瘤作用[J].中华胰腺病学杂志, 2009, 9 (3) :193-197.

[21]魏小雷, 曹俊, 刘文佳, 等.体外负载HSP70-K-ras突变多肽复合物树突细胞的抗胰腺癌作用[J].胃肠病学, 2009, 14 (1) :16-21.

[22]Ashkenazi A, Pai RC, Fong S, et al.Safety and antitumoractivity of recombinant soluble Apo-ligand[J].Clin Invest, 1999, 104 (2) :155-162.

[23]Kim K, Fisher MJ, Xu SQ, et al.Molecular determinants ofresponse to TRAIL in killing of normal and cancer cells[J].Clin Cancer Res, 2000, 6 (2) :335-346.

[24]郭建巍, 秦力维, 蔡美英, 等.肝癌DC疫苗诱导CTL的效应与机理研究[J].免疫学杂志, 2003, 19 (5) :368-371.

[25]Chapoval AI, Tamada K, Chen L, et al.In vitro growth inhi-bition of a broad spectrum of tumor cell lines by activated hu-man dendritic cells[J].Blood, 2000, 95 (7) :2346-2351.

[26]Hirooka Y, Itoh A, Kawashima, et al.A combination therapyof gemcitabine with immunotherapy for patients with inopera-ble locally advanced pancreatic cancer[J].Pancreas, 2009, 38 (3) :e69-74.

[27]Irisawa A, Takagi T, Kanazawa M, et al.Endoscopic ultra-sound-guided fine-needle injection of immature dendriticcells into advanced pancreatic cancer refractory to gemcit-abine:a pilot study[J].Pancreas, 2007, 35 (2) :189-190.

[28]Schmidt T, Ziske C, M rten A, et al.Intratumoral immuniza-tion with tumor RNA-pulsed dendritic cells confers antitu-mor immunity in a C57BL/6 pancreatic murine tumor model[J].Cancer Res, 2003, 63 (24) :8962-8967.

施引文献

资源附件(0)

访问统计