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Preparation of an oral DNA vaccine based on VEGFR2 and investigation of its immune effects in mice
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Abstract:
Abstract Objective: Vascular endothelial growth factor receptor2 plays an important role in tumor growth and metastasis. This study is to prepare an oral VEGFR2 DNA vaccine and to evaluate its immune effects in mice. Methods: The recombinant plasmid pcDNA3.1VR2 targeting the extracellular domains of VEGFR2 was constructed by gene engineering technique. After confirmated by sequencing the pcDNA3.1VR2 vector was transfected into COS7 cells via lipid; the protein expression of VEGFR was identified by Western blotting. Then pcDNA3.1VR2 was transformed into attenuated Salmonella typhimurium SL3261 to develop an oral DNA vaccine against the extracellular domains of VEGFR2. C57BL/6 mice were immunized with the prepared oral vaccine; the serum level of VEGFR2Ag specific antibody was detected by ELISA. The cytotoxic T lymphocyte (CTL) was detected by modified MTT assay. Results：We successfully prepared VEGFR2 DNA vaccine. ELISA results showed that the VEGFR2specific IgG level in oral vaccine group (1.07±0018) was significantly higher than those of the empty vector group (0.14±0.038) and saline group (0.14±0.033)（F=853.17，P=0.000）. The titer of serum antibody increased with the immunization period and the immunization times. As shown by MTT assay, the prepared oral vaccine had specific and efficient cytotoxicity against murine endothelium cells （Ms1） in a dosedependent manner in vitro. The CTL activity in the oral vaccine group (89.38±1.51) was significantly higher than those of the empty vector group (12.99±4.31) and saline group (15.17±2.54) （F=315.42，P=0.000）．Conclusion：We have successfully prepared an oral VEGFR-2 DNA vaccine which can stimulate specific cellular and humoral immune responses in mice, paving a way for further anticancer research.
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Project Supported:
Supported by the Joint Foundation of Science & Technology Bureau of Yunnan Province and Kunming Medical College (No. 2007C0024R)
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