Protective immunity induced by a DNA vaccine cocktail expressing TgSAG1, TgROP2, and the genetic adjuvant HBsAg against Toxoplasma gondii infection.

Toxoplasma gondii is an intracellular obligate parasitic protozoon that can infect all warm-blooded animals, causing zoonotic toxoplasmosis. So far, there is no commercial toxoplasmosis vaccine for human use. In the present study, we constructed a DNA vaccine cocktail which includes the surface protein (SAG1) and the rhoptry protein ROP2 denoted as pEGFP-N1-SAG1-ROP2. In order to improve the efficacy, HBsAg was used as a genetic adjuvant to construct pEGFP-N1-HBsAg-SAG1-ROP2. Two eukaryotic plasmids were transiently transfected into HEK293T cells and the expression was examined using fluorescence microscopy and western blotting. We then immunized Kunming mice intramuscularly with the DNA vaccine. After three immunizations, the immune response was evaluated by measuring antibody levels, cytokine production, percentages of CD4(+) and CD8(+) T lymphocytes, and the survival times of the T. gondii RH strain challenged mice. The results showed that the two DNA vaccines stimulated Th1 responses, and had a higher antibody titer, IL-2, IL-12, and IFN-gamma levels, and percentage of CD4(+) and CD8(+) T lymphocytes than the control group. In addition, mice immunized with the pEGFP-N1-HBsAg-SAG1-ROP2 vaccine showed increased survival times compared with pEGFP-N1-SAG1-ROP2.