Construction of Tumor Vaccine Fused from Dendritic Cells and Walker-256 Cells and Its Effect on Implanted Liver Cancer in Rats_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

WANG Jian , BAI Mingdong , XU Gang.

Department of General Surgery, The Second Affiliated Hospital of Xuzhou Medical College, Xuzhou 221006, Jiangsu Province, China;

Corresponding author：

Keywords：

Dendritic cell; Walker256 cancer cell; Cell fusion; Implanted liver cancer; Immunotherapy; Tumor angiogenesis

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

ObjectiveTo explore the antitumor effect of tumor vaccine fused from dendritic cells (DC) and Walker-256 cancer cells on implanted liver cancer in rats and the related mechanism of inhibition for tumor angiogenesis. MethodsWalker-256 cancer cells and mature DC were fused by 50% polyethylene glycol method for preparation of DC-Walker-256 fusion vaccines. Implanted liver cancer models were established through operations on healthy male SD rats at the age of 6－8 weeks. All the rats were divided into four groups, and rats in each group were injected subcutanely with fusion vaccine (group), mixed cultured cells (group), simple DC (group), and PBS (blank control group), respectively. On 28 d after making model, the rats were put to death, the tumor was observed and pathological essays were prepared. All rats’ spleens were collected and prepared into lymphocyte to detect antigenic specificity cytotoxic T lymphocyte (CTL) by enzymelinked immunosorbent spot (ELISPOT) method. The expressions of VEGF, ANG-1, ANG-2, and MVD were detected by immunohistochemistry. ResultsThe numbers of rats survived in the fusion vaccine group, mixed culture cells group, simple DC group, and blank control group was 8, 5, 6, and 3, respectively. The rats in the other three groups except for fusion vaccine group were manifested as inaction, anorexia, and gloomy fur in some degree as well as ascites. The tumorigenesis was found in all survival rats except for two in the fusion vaccine group. The weight of liver tumors of rats in the fusion vaccine group 〔(32.4±9.2) g〕 was significantly lighter than that in the mixed culture cells group 〔(67.3±5.1) g, P=0.031〕, simple DC group 〔(75.0±8.3) g, P=0.019〕, and blank control group 〔(86.6±10.5) g, P=0.008〕, respectively. The number of tumorspecific CTL of rats in the fusion vaccine group was also significantly higher than that in the other three groups (P=0.019, P=0.025, and P=0.001, respectively). The MVD of tumor tissue in the fusion vaccine group was (24.12±2.32) vessels/HP, which was significantly lower than that in the mixed culture cells group 〔(40.34±1.29) vessels/HP, P=0.025〕, simple DC group 〔(42.36±3.16) vessels/HP, P=0.035〕, and blank control group 〔(56.48±5.16) vessels/HP, P=0.006〕, respectively. The MVD of tumor tissue in the mixed cultured cells group and simple DC group was similar (P=0.165), however, which was significantly lower than that in the blank control group (P=0.040 and P=0.043). The positive rate of VEGFA protein expression was 23.2% in the fusion vaccine group, which was significantly lower than that in the mixed culture cells group (42.5%, P=0.031), simple DC group (61.3%, P=0.019), and blank control group (89.6%, P=0.003), respectively. The positive rate of VEGF-A protein expression in the mixed cultured cells and simple DC groups was similar (P=0.089), however, which was significantly lower than that in the blank control group (P=0.027 and P=0.038). The positive rate of ANG-1 protein expression in the fusion vaccine group (43.2%) was not different from that in the mixed culture cells group (46.3%, P=0.292), simple DC group (51.3%, P=0.183), or blank control group (49.6%, P=0.179), respectively, and the difference of pairwise comparison in latter three groups was not significant (P=0.242, P=0.347, and P=0.182). The positive rate of ANG2 protein expression was 19.2% in the fusion vaccine group, which was significantly lower than that in the mixed culture cells group (62.3%, P=0.007), simple DC group (67.3%, P=0.005), and blank control group (71.6%, P=0.004), respectively, however, the difference of pairwise comparison in latter three groups was not significant (P=0.634, P=0.483, and P=0.379). ConclusionFused vaccine can induce CD8+ CTL aiming at tumor cells and establish the effective antitumor immunity in vivo and also downregulate the level of VEGF and ANG-2 to suppress tumor angiogenesis and thereby achieve the purpose of curing tumor.

Citation： WANG Jian,BAI Mingdong,XU Gang.. Construction of Tumor Vaccine Fused from Dendritic Cells and Walker-256 Cells and Its Effect on Implanted Liver Cancer in Rats. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2011, 18(5): 507-513. doi: Copy

1.	Babatz J, Rllig C, Lbel B, et al. Induction of cellular immune responses against carcinoembryonic antigen in patients with metastatic tumors after vaccination with altered peptide ligandloaded dendritic cells [J]. Cancer Immunol Immunother, 2006, 55(3): 268276.
2.	Nawrocki S, Wysocki PJ, Mackiewicz A. Genetically modified tumour vaccines: an obstacle race to break host tolerance to cancer [J]. Expert Opin Biol Ther, 2001, 1(2): 193204.
3.	Langley RR, Ramirez KM, Tsan RZ, et al. Tissuespecific microvascular endothelial cell lines from H2K(b)tsA58 mice for studies of angiogenesis and metastasis [J]. Cancer Res, 2003, 63(11): 29712976.
4.	St Croix B, Rago C, Velculescu V, et al. Genes expressed in human tumor endothelium [J]. Science, 2000, 289(5482): 11971202.
5.	An FQ, Matsuda M, Fujii H, et al. Expression of vascular endothelial growth factor in surgical specimens of hepatocellular carcinoma [J]. J Cancer Res Clin Oncol, 2000, 126(3): 153160.
6.	Moon WS, Rhyu KH, Kang MJ, et al. Overexpression of VEGF and angiopoietin 2: a key to high vascularity of hepatocellular carcinoma? [J]. Mod Pathol, 2003, 16(6): 552557.
7.	Tanaka S, Mori M, Sakamoto Y, et al. Biologic significance of angiopoietin2 expression in human hepatocellular carcinoma [J]. J Clin Invest, 1999, 103(3): 341345.
8.	Mitsuhashi N, Shimizu H, Ohtsuka M, et al. Angiopoietins and Tie2 expression in angiogenesis and proliferation of human hepatocellular carcinoma [J]. Hepatology, 2003, 37(5): 11051113.
9.	Gong J, Chen D, Kashiwaba M, et al. Induction of antitumor activity by immunization with fusions of dendritic cell and carcinoma cells [J]. Nat Med, 1997, 3(5): 558561.
10.	Sun K, Wang L, Zhang Y. Dendritic cell as therapeutic vaccines against tumors and its role in therapy for hepatocellular carcinoma [J]. Cell Mol Immunol, 2006, 3(3): 197203.
11.	Guo Y, Wu M, Chen H, et al. Effective tumor vaccine generated by fusion of hepatoma cells with activated B cells [J]. Science, 1994, 263(5146): 518520.
12.	谭广, 任金帅, 王忠裕, 等. TNFα增强树突状细胞诱导的抗胰腺癌免疫应答的实验研究 [J]. 中国普外基础与临床杂志, 2006, 13(6): 689692.
13.	薛刚, 曹永宽, 王培红, 等. 肿瘤细胞裂解物致敏的树突状细胞对结肠癌LoVo细胞的杀伤作用 [J]. 中国普外基础与临床杂志, 2009, 16(1): 4851.
14.	Hildenbrand B, Sauer B, Kalis O, et al. Immunotherapy of patients with hormonerefractory prostate carcinoma pretreated with interferongamma and vaccinated with autologous PSApeptide loaded dendritic cells-a pilot study [J]. Prostate, 2007, 67(5): 500508.
15.	Venier C, Guthmann MD, Fernández LE, et al. Innateimmunity cytokines induced by very small size proteoliposomes, a Neisseriaderived immunological adjuvant [J]. Clin Exp Immunol, 2007, 147(2): 379388.
16.	Folkman J. Role of angiogenesis in tumor growth and metastasis [J]. Semin Oncol, 2002, 29(6 Suppl 16): 1518.
17.	Foote RL, Weidner N, Harris J, et al. Evaluation of tumor angiogenesis measured with microvessel density (MVD) as a prognostic indicator in nasopharyngeal carcinoma: results of RTOG 9505 [J]. Int J Radiat Oncol Biol Phys, 2005, 61(3): 745753.
18.	Folkman J. Angiogenesis: an organizing principle for drug discovery? [J]. Nat Rev Drug Discov, 2007, 6(4): 273286.
19.	Poon RT, Ng IO, Lau C, et al. Tumor microvessel density as a predictor of recurrence after resection of hepatocellular carcinoma: a prospective study [J]. J Clin Oncol, 2002, 20(7): 17751785.
20.	Shibuya M, ClaessonWelsh L. Signal transduction by VEGF receptors in regulation of angiogenesis and lymphangiogenesis [J]. Exp Cell Res, 2006, 312(5): 549560.
21.	魏学明, 顾国利, 任力, 等. 胃癌组织中EGFR、CerbB2、VEGF及COX2的表达及意义 [J] . 中国普外基础与临床杂志, 2009, 16(11): 900905.
22.	Paschke M. Phage display systems and their applications [J]. Appl Microliol Biotechnol, 2006, 70(1): 211.
23.	Mi D, Yi J, Liu E, et al. Relationship between PTEN and VEGF expression and clinicopathological characteristics in HCC [J]. J Huazhong Univ Sci Technolog Med Sci, 2006, 26(6): 682685.
24.	Kanematsu M, Osada S, Amaoka N, et al. Expression of vascular endothelial growth factor in hepatocellular carcinoma and the surrounding liver: correlation with angiographically assisted CT [J]. AJR Am J Roentgenol, 2004, 183(6): 15851593.

1. Babatz J, Rllig C, Lbel B, et al. Induction of cellular immune responses against carcinoembryonic antigen in patients with metastatic tumors after vaccination with altered peptide ligandloaded dendritic cells [J]. Cancer Immunol Immunother, 2006, 55(3): 268276.
2. Nawrocki S, Wysocki PJ, Mackiewicz A. Genetically modified tumour vaccines: an obstacle race to break host tolerance to cancer [J]. Expert Opin Biol Ther, 2001, 1(2): 193204.
3. Langley RR, Ramirez KM, Tsan RZ, et al. Tissuespecific microvascular endothelial cell lines from H2K(b)tsA58 mice for studies of angiogenesis and metastasis [J]. Cancer Res, 2003, 63(11): 29712976.
4. St Croix B, Rago C, Velculescu V, et al. Genes expressed in human tumor endothelium [J]. Science, 2000, 289(5482): 11971202.
5. An FQ, Matsuda M, Fujii H, et al. Expression of vascular endothelial growth factor in surgical specimens of hepatocellular carcinoma [J]. J Cancer Res Clin Oncol, 2000, 126(3): 153160.
6. Moon WS, Rhyu KH, Kang MJ, et al. Overexpression of VEGF and angiopoietin 2: a key to high vascularity of hepatocellular carcinoma? [J]. Mod Pathol, 2003, 16(6): 552557.
7. Tanaka S, Mori M, Sakamoto Y, et al. Biologic significance of angiopoietin2 expression in human hepatocellular carcinoma [J]. J Clin Invest, 1999, 103(3): 341345.
8. Mitsuhashi N, Shimizu H, Ohtsuka M, et al. Angiopoietins and Tie2 expression in angiogenesis and proliferation of human hepatocellular carcinoma [J]. Hepatology, 2003, 37(5): 11051113.
9. Gong J, Chen D, Kashiwaba M, et al. Induction of antitumor activity by immunization with fusions of dendritic cell and carcinoma cells [J]. Nat Med, 1997, 3(5): 558561.
10. Sun K, Wang L, Zhang Y. Dendritic cell as therapeutic vaccines against tumors and its role in therapy for hepatocellular carcinoma [J]. Cell Mol Immunol, 2006, 3(3): 197203.
11. Guo Y, Wu M, Chen H, et al. Effective tumor vaccine generated by fusion of hepatoma cells with activated B cells [J]. Science, 1994, 263(5146): 518520.
12. 谭广, 任金帅, 王忠裕, 等. TNFα增强树突状细胞诱导的抗胰腺癌免疫应答的实验研究 [J]. 中国普外基础与临床杂志, 2006, 13(6): 689692.
13. 薛刚, 曹永宽, 王培红, 等. 肿瘤细胞裂解物致敏的树突状细胞对结肠癌LoVo细胞的杀伤作用 [J]. 中国普外基础与临床杂志, 2009, 16(1): 4851.
14. Hildenbrand B, Sauer B, Kalis O, et al. Immunotherapy of patients with hormonerefractory prostate carcinoma pretreated with interferongamma and vaccinated with autologous PSApeptide loaded dendritic cells-a pilot study [J]. Prostate, 2007, 67(5): 500508.
15. Venier C, Guthmann MD, Fernández LE, et al. Innateimmunity cytokines induced by very small size proteoliposomes, a Neisseriaderived immunological adjuvant [J]. Clin Exp Immunol, 2007, 147(2): 379388.
16. Folkman J. Role of angiogenesis in tumor growth and metastasis [J]. Semin Oncol, 2002, 29(6 Suppl 16): 1518.
17. Foote RL, Weidner N, Harris J, et al. Evaluation of tumor angiogenesis measured with microvessel density (MVD) as a prognostic indicator in nasopharyngeal carcinoma: results of RTOG 9505 [J]. Int J Radiat Oncol Biol Phys, 2005, 61(3): 745753.
18. Folkman J. Angiogenesis: an organizing principle for drug discovery? [J]. Nat Rev Drug Discov, 2007, 6(4): 273286.
19. Poon RT, Ng IO, Lau C, et al. Tumor microvessel density as a predictor of recurrence after resection of hepatocellular carcinoma: a prospective study [J]. J Clin Oncol, 2002, 20(7): 17751785.
20. Shibuya M, ClaessonWelsh L. Signal transduction by VEGF receptors in regulation of angiogenesis and lymphangiogenesis [J]. Exp Cell Res, 2006, 312(5): 549560.
21. 魏学明, 顾国利, 任力, 等. 胃癌组织中EGFR、CerbB2、VEGF及COX2的表达及意义 [J] . 中国普外基础与临床杂志, 2009, 16(11): 900905.
22. Paschke M. Phage display systems and their applications [J]. Appl Microliol Biotechnol, 2006, 70(1): 211.
23. Mi D, Yi J, Liu E, et al. Relationship between PTEN and VEGF expression and clinicopathological characteristics in HCC [J]. J Huazhong Univ Sci Technolog Med Sci, 2006, 26(6): 682685.
24. Kanematsu M, Osada S, Amaoka N, et al. Expression of vascular endothelial growth factor in hepatocellular carcinoma and the surrounding liver: correlation with angiographically assisted CT [J]. AJR Am J Roentgenol, 2004, 183(6): 15851593.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Construction of Tumor Vaccine Fused from Dendritic Cells and Walker-256 Cells and Its Effect on Implanted Liver Cancer in Rats

Abstract Full text Figures/Tables Video References Cited by

Format

Content