Flowjo serial number8/14/2023 The avirulent RHΔ ompdcΔ uprt strain can provide strong anti- T. In addition, compared with nonimmunized cats, cats immunized with the mutant produced high levels of antibodies and cytokines ( P < 0.05), and notably decreased the shedding numbers of oocysts in feces (95.3%). The immunized sera and splenocytes, especially CD8 + T cells, could significantly extend ( P < 0.05) the survival time of mice challenged with the RHΔ ku80 strain compared with naïve mice. Remarkably, all RHΔ ompdcΔ uprt-vaccinated mice survived a lethal challenge with RHΔ ku80 and ME49 and WH6 strains. Furthermore, compared with nonimmunized group, high levels of IgG (IgG1 and IgG2a) antibodies and cytokines (IFN-γ, IL-4, IL-10, IL-2 and IL-12) in mice were detected by the mutant ( P < 0.05). Notably, limited pathological changes were found in tissues from RHΔ ompdcΔ uprt-injected mice. In addition, the mutant exhibited virulence attenuation in both murine (BALB/c and BALB/c-nu) and cat models. Compared with the wild-type strain, the mutant notably reduced proliferation ( P < 0.05). The RHΔ ompdcΔ uprt were constructed by the CRISPR-Cas9 system. GraphPad Prism software was used to conduct the log-rank (Mantel–Cox) test, Student’s t test and one-way ANOVA. Furthermore, to discover the effective immune element against toxoplasmosis, passive immunizations were carried out. Finally, the immunoprotective effects were evaluated by challenge with tachyzoites of different strains in mice or cysts of the ME49 strain in cats. Subsequently, the immune responses induced by this mutant in mice and cats were detected, including antibody titers, cytokine levels, and subsets of T lymphocytes. Then, the intracellular proliferation and virulence of this mutant strain were evaluated. gondii were deleted through the CRISPR-Cas9 system. Methodsįirst, the ompdc and uprt genes of T. gondii vaccine for the control of toxoplasmosis, and this study is to explore the immunoprotective effects of a live attenuated vaccine in mice and cats. It is essential to develop an effective anti- T. Toxoplasma gondii is an obligate intracellular apicomplexan parasite and is responsible for zoonotic toxoplasmosis.
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