| Objective:Study the immune response and immune protective effect of Toxoplasma gondii recombinant protein vaccine based on molecular adjuvant EDA,FliC and TAT and Toxoplasma gondii SAG1 antigen in mice.Methods:Firstly,we constructed four prokaryotic expression vector:pET-28a-EDA-SAG1,pET-28a-FliC-SAG1,pET-28a-TAT-SAG1 and pET-28a-SAGl;then the expression vector were transformed into E.coli for the protein expressiong.Finally,SDS-PAGE and Western-blot were used to determine the expression of the recombinant protein.Mice were immunized with the recombinant protein to detect the humoral immune response and cellular immune response.Then the mice were infected with Toxoplasma gondii RH strain,and survival rate of mice were calculated to study the immunoprotective effect of recombinant protein vaccines.Results:1.Four prokaryotic expression vector were successful constructed.2.SDS-PAGE and Western-blot results showed that the proteins molecular weight size of rSAGl is about 28 kDa,rTAT-SAG1 is about 29 kDa,rEDA-SAG1 is about 38 kDa,and rFliC-SAG1 is about 58 kDa;3.After the mice were immunized with the recombinant proteins,there are significant differences between the molecular adjuvant groups and control group in the humoral immune response.For the cell proliferation,EDA-SAG1 group had the highest proliferation rate than other groups.For the cytokines expression,the molecular adjuvant groups produced more IFN-y than Freund’s adjuvant group and the control group.TAT-SAG 1 group produce more IL-12 than other groups.And EDA-SAG1 group produce more IL-2 than other groups.And there are significant differences for IL-10 production between TAT-SAG1 group and Freund’s adjuvant group.After the mice were challenged with Toxoplasma gondii RH strain,the molecular adjuvant EDA-SAG1 group had the highest survival rate and mice in this group survived three days longer than the control group.Conclusion:The molecular adjuvant EDA,FliC,and TAT can significantly enhance the immune response of Toxoplasma gondii recombinant protein vaccine.What’s more,they can also improve the immune protection. |
