Investigation on the anti-tumor efficacy by expression of GPI-anchored mIL-21 on the surface of B16F10 cells in C57BL/6 mice.
GPI-anchored membrane cytokines have been shown to play an important role in host immune response against tumor cells. In the present study, we constructed the tumor vaccine expressing mIL-21 in the GPI-anchored form and investigated its anti-tumor effect in C57BL/6 mice model. The fusion genes containing mIL-21 and the GPI anchor signal sequence was acquired by overlaping PCR, inserted into plasmid pcDNA3.1 to form the pcDNA3.1 mIL-21-GPI recombinant, which was transfected into the B16F10 cells, and the tumor vaccine based on B16F10 cells expressing the GPI-anchored membrane mIL-21 was generated. Through transfection, it was found that GPI-anchored membrane mIL-21 has no proliferate impact on B16F10 cells, but it was functional and reflected in inducing CD3-activated murine splenocytes proliferation response to B16F10 cells, improving the cytotoxicities of CTL and NK cells, increasing the numbers of splenocytes-producing IFN-gamma in mice, augmenting therapeutic effect of tumor and prolonging longevity effects in tumor-bearing mice injected with the inactivated GPI-anchored mIL-21 tumor vaccine. We concluded the expression of mIL-21 on the B16F10 cells surface in the GPI-anchored form was proved to be effective in activating immune responses against tumor cells, and our results provided a good foundation for further investigating the immunotherapy of tumor by GPI-mIL-21.
Zhao
Fengshu
F
Department of Pathogenic Biology and Immunology, School of Basic Medical Science, Southeast University, Nanjing, China.
Dou
Jun
J
Wang
Jing
J
Chu
Lili
L
Tang
Quan
Q
Wang
Yongfang
Y
Li
Yating
Y
Cao
Minggang
M
Hu
Weihua
W
Hu
Kai
K
Feng He
Xiang
X
Gu
Ning
N
D000818:Animals; D000906:Antibodies / Q000276:immunology; D017252:CD3 Complex / Q000276:immunology; D019496:Cancer Vaccines / Q000235:genetics / Q000276:immunology* / Q000627:therapeutic use; D045744:Cell Line, Tumor; D050939:Gene Fusion; D005822:Genetic Vectors; D017261:Glycosylphosphatidylinositols / Q000235:genetics / Q000276:immunology*; D016233:Immunotherapy, Active; D007378:Interleukins / Q000235:genetics / Q000276:immunology*; D007694:Killer Cells, Natural / Q000276:immunology; D008213:Lymphocyte Activation; D008546:Melanoma, Experimental / Q000276:immunology / Q000628:therapy*; D051379:Mice; D008810:Mice, Inbred C57BL; D012878:Skin Neoplasms / Q000276:immunology / Q000628:therapy*; D013154:Spleen / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014162:Transfection; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology* / Q000627:therapeutic use; D000098546:Interleukin-21
D000906:Antibodies; D017252:CD3 Complex; D019496:Cancer Vaccines; D017261:Glycosylphosphatidylinositols; D007378:Interleukins; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7; D019444:Vaccines, DNA; D000098546:Interleukin-21