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Antitumor effect of recombinant cEGFRrFc infusion DNA vaccine against transplanted melanoma in mice
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Abstract:
Abstract Objective: To construct the recombinant DNA vaccine by fusing cEGFR with IgG Fc, and to observe the inhibitory effect of the constructed DNA vaccine against the transplanted melanoma in mice. Methods: The extracellular domain of xenogeneic chicken epidermal growth factor receptor (cEGFR) was fused with rabbit IgG Fc; and the product was insert into PVAX1 eukaryotic expression vector to construct the recombinant pVAX1/cEGFRrFc DNA vaccine. Expression of the fusion protein in melanoma B16 cells was detected by immunofluorescent assay after transfection. In vivo expression of the fusion protein in immunized C57BL/6J mice was detected by Western blotting assay. The DNA vaccines were used to immunize C57 mice 3 times before transplantation of B16 melanoma cells . The weights of tumors were determined after 14 d to calculate the antitumor rate and the survival rate of the mice was observed. The serum level of the specific antibody against human EGFR was determined by ELISA. Subgroups of spleen T lymphocytes were examined by FACS. Results: The eukaryotic expression vector pVAX1/ cEGFRrFc was successfully constructed. Effective expression of fusion protein was detectable in B16 cells after transfection with recombinant plasmid and stable expression was detected in mice after vaccine immunization. The fused DNA vaccine inhibited the growth of transplanted melanoma, with the inhibitory rate being 54% (P<0.01). The vaccine also prolonged the survival period of the mice transplanted with B16 cancer cells (the survival rate was 40% 30 d after inoculation). The vaccine also induced production of specific antibody against human EGFR (1∶1 000) and T cell immunoresponse (The CD8+ subgroups of spleen T lymphocyte were significantly increased \[P<0.01\]). Conclusions: The DNA vaccine containing cEGFR and IgG Fc is an effective antitumor vaccine against EGFRpositive tumor.
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Supported by the Key Science and Technology Research Project of Shanghai
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