Compound DNA vaccine encoding SAG1/ SAG3 with A(2)/B subunit of cholera toxin as a genetic adjuvant protects BALB/c mice against Toxoplasma gondii

BACKGROUND: Intracellular parasites, such as T. gondii, present a plurality of antigens because of the complexity of its life cycle. Compound DNA vaccines bring a new approach and hope for the treatment of toxoplasmosis. In this study, a DNA vaccine encoding two major surface antigens SAG1, SAG3 from T. gondii, with A(2)/B subunit of cholera toxin as a genetic adjuvant was constructed. METHODS: BALB/c mice were immunized intramuscularly with PBS, pcDNA3.1, pSAG1, pSAG1/SAG3 and pSAG1/SAG3-CTXA(2)/B three times separately. Immunized mice were tested for IgG antibody and IFN-γ and IL-4 production by ELISA. The proliferation of T cells was measured by DNA synthesis assay and the lymphocyte subsets of spleen cells by flow cytometry. All the immunized mice were challenged with 10(3) highly virulent RH tachyzoites of Toxoplasma gondii intraperitoneally and the survival times were recorded. RESULTS: An enhanced production of IgG antibodies, antigen-specific lymphocyte proliferation and IFN-γ production from splenic cells were induced in mice immunized with pSAG1/SAG3 compared to mice immunized with pSAG1 (P<0.05). Introduction of CTXA(2)/B further enhanced the Th1 cell-mediated immunity with higher levels of IFN-γ, lymphocyte proliferation activity and percentage of CD8(+) T-cells. When challenged with lethal doses of T. gondii (1×10(3)), all control mice (PBS and empty plasmid group) died within 6 days. Mice immunized with pSAG1 died within 8 days. While 20% and 40% survival rate were achieved from mice immunized with pSAG1/SAG3 and pSAG1/SAG3-CTXA(2)/B. CONCLUSIONS: This study indicates the compound DNA vaccine encoding T. gondii antigens SAG1, SAG3 with CTXA(2)/B gene was a promising DNA vaccine candidate against toxoplasmosis, which could effectively enhance the humoral and cellular immune response and prolong survival time in vaccinated mice.