Construction of DNA vaccine encoding murine AFP and its specific anti-tumor effect on C57BL/6 mice / 第三军医大学学报

ABSTRACT

Objective To clone the murine ?-fetoprotein gene,construct the DNA vaccine of mouse AFP and to observe the specific cellular immunologic responses and the anti-tumor responses in mice induced by this vaccine.Methods The murine AFP gene was amplified by RT-PCR from total RNA extracted from Hepa1-6 cells and cloned into the vector pcDNA3.1 to construct pmAFP.The DNA vaccine was identified by restriction enzyme analysis,sequencing and expression.EL-4(mAFP) was developed by stable transfection of EL-4 cells with pmAFP.Mice immunized with DNA vaccine by intramuscular injection were inoculated with EL-4(mAFP) cells at back to observe the inhibitory effect of the immunization on tumor.The frequency of cells producing IFN-? in splenocytes harvested from the immunized mice was measured by ELISPOT.Blood samples were collected from the immunized mice to check the functions of liver and kidney.Results The murine AFP gene was successfully cloned by RT-PCR.DNA vaccine was successfully constructed.The expression of mAFP mRNA in EL-4(mAFP) was confirmed by RT-PCR.The tumor volume in pmAFP vaccine group(1 042.42?123.71 mm~(3)) was significantly smaller than that of other groups(P