• Vaccine Name: TgMIC1-4-6
• Target Pathogen: Toxoplasma gondii
• Target Disease: Toxoplasmosis
• Type: Subunit vaccine
• Status: Research
• Host Species for Licensed Use: Human
• Antigen: MIC1, MIC4, and MIC6 (Pinzan et al., 2015)
• MIC6 gene engineering:
  • Type: Recombinant protein preparation
  • Description: The proteins were expressed by E. coli BL21 (DE3) (Novagen) cells transformed with pET21b-MIC6. (Pinzan et al., 2015)
  • Detailed Gene Information: Click Here.
• MIC1 gene engineering:
  • Type: Recombinant protein preparation
  • Description: MIC1 was PCR amplified and was cloned into pDONR201 vector and yielded pENTR-TgMIC1. Inserts from pENTR-TgMIC1 were transferred into pDEST17 vectors and yielded pEXP17-TgMIC1. The proteins were expressed by E. coli BL21 (DE3) (Novagen) cells transformed with pDEST17-MIC1. (Pinzan et al., 2015)
  • Detailed Gene Information: Click Here.
• MIC4 gene engineering:
  • Type: Recombinant protein preparation
  • Description: MIC4 was PCR amplified and was cloned into pDONR201 vector and yielded pENTR-TgMIC4. Inserts from pENTR-TgMIC4 were transferred into pDEST17 vectors and yielded pEXP17-TgMIC4. The proteins were expressed by E. coli BL21 (DE3) (Novagen) cells transformed with pDEST17-MIC4. (Pinzan et al., 2015)
  • Detailed Gene Information: Click Here.
• Immunization Route: subcutaneous injection
• Description: T. gondii subunit vaccine expressing MIC1, MIC4, and MIC6 (Pinzan et al., 2015)

Mouse Response

• Host Strain: C57BL/6 (Pinzan et al., 2015)
• Host age: 6–7 weeks (Pinzan et al., 2015)
• Vaccination Protocol: Mice were subcutaneously (s.c.) injected with TgMIC1 (10 μg), TgMIC4 (10μg), TgMIC6 (10μg), TgMIC1-4 (5 μg of each protein), TgMIC1-4-6 (3.3μg of each protein), or lactose-binding proteins (Lac+, 10 μg) emulsified in Freund’s complete adjuvant. Animals were boosted at the same dose and regimen on day 15 and 30 after first injection, now emulsified in Freund’s incomplete adjuvant. A control group was injected at the same regimen with PBS emulsified in Freund’s adjuvant (vehicle). (Pinzan et al., 2015)
• Challenge Protocol: One month after the last immunization procedure, the mice were orally infected with 80 cysts of the ME49 strain and were monitored and recorded for 30 days to compute the survival rates. The brain of the mice infected with 40 cysts was removed 1 month after the challenge and the mean number of cysts per brain was determined. (Pinzan et al., 2015)
• Efficacy: Mice immunized with TgMIC1-4-6, compared with non-immunized mice, showed reductions in brain cysts by 67.8%. Non-immunized control mice started dying 6 days after T. gondii infection, and they were all dead by day 11 post infection. Effective and highly significant protection was demonstrated in mice immunized with TgMIC1-4-6, since 80% of the mice survived to the acute phase of infection. (Pinzan et al., 2015)