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Chinese Journal of Clinical Pharmacology and Therapeutics ›› 2021, Vol. 26 ›› Issue (1): 24-29.doi: 10.12092/j.issn.1009-2501.2021.01.004

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Preparation and the effect of antitumor of DNA plasmid lipidosome vaccine based VEGFR2 extracellular region by immunization activated in vitro

CHEN Minfang1, CHEN Lihua3, XIE Liyun2, XU Fenfen2, XIA Aixiao2, LIN Zhong2   

  1. 1 Department of Thoracic Surgery, Wenling Traditional Chinese Medicine Hospital, Taizhou 317500, Zhejiang, China
  • Received:2020-04-08 Revised:2020-09-22 Online:2021-01-26 Published:2021-02-01

Abstract: AIM: The DNA plasmid lipidosome (LP) vaccine based VEGFR2 extracellular region (exVEGFR2) was prepared in order to provide a new approach for cancer active immunotherapy.  METHODS: High fidelity PCR was used to amplify the target sequence of exVEGFR2 with two restriction site of KpnⅠ and XbaⅠ. The plasmid of pCMV/exVEGFR2 was constructed by connected exVEGFR2 with pCMV empty plasmid. The activity of immune activation was detected by ELISA. CTLs mediated cytotoxic activity was analyzed by 51Cr release assay. RESULTS: The 90 000 target specific band of VEGFR2 was detected by Western blot. After 6 weeks since the first time vaccination, an intense specific immune response of anti-VEGFR2 was detected by ELISA in the serum from the C57BL/6 mouse vaccinated with LP-pCMV/exVEGFR2 vaccine. The T cells from the spleen of mouse immunized with the vaccine induced the cytotoxicity effect on CT-26 with VEGFR2 positive. CONCLUSION: The results of the specific immune response of anti-VEGFR2 in vivo and the antitumor cytotoxicity in vitro by vaccinated with LP-pCMV/exVEGFR2 vaccine in mouse model, would lay the foundation for further study of VEGFR2 positive tumor treated in vivo by the active immunotherapy.

Key words: VEGFR2, lipidosome, pCMV plasmid, vaccine

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