Assessment of the immunogenicity and protective efficiency of a novel dual-promoter DNA vaccine, harboring SAG1 and GRA7 genes, from RH strain of Toxoplasma gondii in BALB/c mice

Abstract

Background

Toxoplasmosis, a protozoan parasitic disease caused by Toxoplasma gondii, has been a serious human and veterinary medicine problem with global distribution. In the current study, we assessed immunogenicity and protective efficiency of a novel dual-promoter DNA vaccine, harboring SAG1 and GRA7 genes, from RH strain of Toxoplasma gondii (T. gondii) with or without CpG-ODN as adjuvant in a murine model.

Methods

BALB/c mice were immunized intramuscularly with pVitro-SAG1-GRA7 alone and pVitro-SAG1-GRA7 with CpG-ODN three times at three-week intervals. Enzyme-linked immunosorbent assay (ELISA) was used to assess total IgG, IgG1 and IgG2a antibodies and gamma interferon (IFN-γ) and interleukin-10 (IL-10) cytokines in mice sera. Four weeks post final vaccination, MTT assay and lethal challenge-infection with 1×10³ tachyzoites of T. gondii RH strain were carried out to assess stimulation index (SI) and mice survival time, respectively.

Results

The IgG levels in mice immunized with multicomponent vaccines, including pVitro-SAG1–GRA7 alone and pVitro-SAG1–GRA7 with CpG-ODN, were significantly higher than those in control mice or single-gene DNA-vaccinated ones (P<0.05). Furthermore, level of IgG2a in mice receiving pVitro-SAG1–GRA7 with CpG-ODN was significantly higher than that in mice receiving pVitro-SAG1-GRA7 alone (P<0.05). The Toxoplasma lysate antigen (TLA)-stimulated lymphocytes from pVitro-SAG1-GRA7 with CpG-ODN group responded more dramatically than those from control groups or single-gene DNA-vaccinated groups (P<0.001). The pVitro-SAG1-GRA7 with CpG-ODN-vaccinated mice developed high levels of IgG2a and IFN-γ (P<0.001) and low levels of IgG1 and IL-10, compared to control groups, suggesting a modulated immune response type Th1. In addition, survival time of the mice immunized with pVitro-SAG1-GRA7 with CpG-ODN was significantly extended, compared to controls (P<0.05); however, all mice died.

Conclusion

The multivalent pVitro-SAG1-GRA7 DNA vaccine with CpG-ODN adjuvant is a promising vaccine candidate against toxoplasmosis.

Keywords:

• Toxoplasma gondii
• RH strain
• SAG1
• GRA7
• pVitro2-neo-mcs
• DNA vaccine
• BALB/c mice
• CpG-ODN adjuvant

Acknowledgments

This study was supported by the Tehran University of Medical Sciences and Health Services (Project No. 21648), Tehran, Iran. We would like to acknowledge all staff within the Toxoplasmosis Laboratory, Department of Medical Parasitology and Mycology, Tehran University of Medical Sciences, for their useful help.

Disclosure

The authors report no conflicts of interest in this work.