Genetic Disruption of Toxoplasma gondii peroxiredoxin (TgPrx) 1 and 3 Reveals the Essential Role of TgPrx3 in Protecting Mice from Fatal Consequences of Toxoplasmosis

• Published in: International journal of molecular sciences Vol. 23; no. 6; p. 3076
• Main Authors: Fereig, Ragab M, Nishikawa, Yoshifumi
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 12.03.2022; MDPI
• Subjects: Animals; Cloning; CRISPR; Economic impact; Egress; Female; Genes; Immune response; Immunology; Infections; Interferon; Interleukin 12; Interleukin 6; Localization; Lymphocytes; Macrophages; Male; Mice; Mice, Inbred C57BL; Mice, Inbred ICR; Mice, SCID; Mitochondria; Monoclonal antibodies; Parasites; Peroxiredoxin; Peroxiredoxins - genetics; Proteins; Protozoan Proteins - genetics; Tachyzoites; Toxoplasma - genetics; Toxoplasma gondii; Toxoplasmosis; Toxoplasmosis - parasitology; Vaccines; virulence; Western blotting; γ-Interferon; peroxiredoxin; Toxoplasma gondii; virulence; toxoplasmosis
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms23063076

is a worldwide protozoan parasite that endangers human health and causes enormous economic losses to the animal production sector. A safe and effective vaccine or treatment is needed to reduce these hazards. In this study, we revealed the cyto-nuclear and mitochondrial localization of TgPrx1 and TgPrx3 proteins, respectively. We knocked out the ( KO) 1 and 3 genes using a parental type II Prugniaud strain lacking KU80 and HXGPRT genes (PruΔku80Δhxgprt) via CRISPR-Cas9 technology. The successful KO was confirmed using PCR, IFAT, and Western blotting in two clones of both target genes, named KO and KO. Regarding assays, no significant variations between any of the knocked-out clones in KO or KO parasite strains, or even PruΔku80Δhxgprt, were obtained in rates of infection, proliferation, or egress. Nevertheless, mice that were infected with tachyzoites of the KO strain showed a marked decrease in survival rate compared with KO- and PruΔku80Δhxgprt-infected mice. This effect was confirmed using different mouse strains (ICR and C57BL/6J mice), sexes (male and female), and immunological backgrounds (ICR and SCID mice). In addition, KO and KO induced high levels of interferon gamma (IFN-γ) in infected mice at 8 days post infection, and increased IL-6 and IL-12p40 production from murine macrophages cultivated . The results of the present study suggested that can induce anti- immune responses that protect the mice from fatal consequences of toxoplasmosis. The results of our current and previous studies represent as an excellent candidate for sub-unit vaccines, suggesting it may contribute to the control of toxoplasmosis for susceptible humans and animals.