Combining DNA Vaccine And Protein Vaccine Based On Porcine Endoglin Encoding The Extracellular Domain Mediates Anti-Tumor Effects In Mouse Models

Objective: The current studies explored the anti-tumor effects by using combination of recombined porcine endoglin DNA vaccine and protein vaccine to study whether the combinational vaccine was better than only one of them in mouse models,and investigated preliminary the mechanism of angiogenesis.Methods: Restrict endonuclease response and agarose gel electrophoresis were used to recognize the construct and molecular of the recombined porcine endoglin plasmid, and the plasmid was confirmed to be correctly expressed in eukaryotic COS-1 cell in vitro. Then we planted CT26 cells in subskin of the right arm and prepared the recombinant as DNA vaccine and the expressed production of the plasmid as protein vaccine. We injected the DNA vaccine and or protein vaccine in the subskin or the muscle of tumor mice. In the study of anti-tumor response, we classed the mice into four groups and the fist group was treated with physiological saline, the second and the third were treated with DNA vaccine and protein vaccine in single, the last group was treated with both. And we respectively named them as NS,DV,PV and CV group. Measured the diameter of the tumor ,calculated the tumor volume with the formula(v=0.52×length×wideth) and recorded the survival time of the tumor mice. Furthermore, anti-CD31 antibody was used to survey microvessel density of the tumor vascular by immunehistochemistry. In addition,we used the same number of mice grouped as the anti-tumor effect observation in order to examine the side effects in therapy,but tumor cells were not injected.Two weeks after four times immunization,mice were put partly to death and the blood samles were collected,and the spleen organs were taken out of. The other mice remained live for seven weeks. During these days we intensely paid attention to the change of mice in skin,appetite ,action and so on. In addtion,we examined the routine of blood,creatinine,GOT and GPT to determine the state of the hepar and kidney. After that, Western blot,ELISPOT assay and ELISA were used to indicate the affairs of antibody. The function of CTL was proved by the release of 51Cr which can display the mechanism of anti-tumor used DNA vaccine and protein vaccine.Result:1,The current study showed the right segments just as prediction by restrict endonuclease response and the eukaryotic plasmid encoding endoglin correctly expressed in...