Anti-Bladder Tumor Effect Of Ag85A DNA Vaccine And Ag85B DNA Vaccine On Tumor-bearing Mice

Objective:To study the anti-bladder tumor effect of Ag85A DNA vaccine and Ag85B DNA vaccine on tumor-bearing mice.Methods:pcDNA3.1-Ag85A plasmids and pcDNA3.1-Ag85B plasmids were extracted from E.coli JM109.The digested plasmids were analyzed by agarose gel electrophoresis.BTT739 cells (1×10~6 cells)were inoculated subcutaneously into the left posterior limb of the mice.We classed the mice into the five groups.The first group was treated with physiological saline,the second was treated with pcDNA3.1,the third was treated with BCG,the fourth and the last group was treated with Ag85A DNA vaccine and Ag85B DNA vaccine in single.The tumor volume was measured for a long time.Calculated the tumor volume with the formula(V=0.52×L×W~2). We intensely paid attention to the change of mice in skin,appetite,action and so on.28 days after the inoculation,the mice were killed and the tumors were removed to study the tumor weight.And the tumor inhibition rate was calculated.The tumors were observed their pathological changes.Spleen was removed aseptically.The percentages of CD4~+T cell and CD8~+T cell were measured by performing flow cytometry.In addition,IFN-γin sera of the tumor-bearing mice were assayed with indirect enzyme-linked immuno-absordent assay (ELISA).Result:(1)pcDNA3.1-Ag85A plasmids and pcDNA3.1-Ag85B plasmids were extracted from E.coli JM109.(2)In the mice model with pre-established subcutaneous BTT739 bladder tumor,Ag85B DNA vaccine could inhibit tumor growth,but the immunization of the BCG inhibited the tumor growth most significantly when compared with Ag85B DNA vaccine.(3)Histological examination showed that obvious tumor necrosis and infiltration of inflammatory cells was present inside the tumors of the tumor-bearing mice immunized with Ag85B DNA vaccine.(4)The result of the percentage of CD4~+T cell,CD8~+T cell and the relative value of CD4~+/CD8~+ showed that there was significant difference between the Ag85B DNA vaccine group and the control group(P＜0.01),there was no significant difference between the Ag85A DNA vaccine group and the control group(P＞0.05).(5)Immunization of mice with Ag85B DNA vaccine and Ag85A DNA vaccine could increase the production of IFN-γ.Conclusion:(1)The Ag85B DNA vaccine could induce strong Thl immune response and elicit significant anti-bladder tumor effect.(2)The Ag85A DNA vaccine may not induce the effective anti-bladder tumor effect.