Toxoplasma gondii, a specialized intracellular parasite, causes a widespread zoonotic disease and is a severe threat to social and economic development.There is a lack of effective drugs and vaccines against T. gondii infection.Recently, mRNA vaccines have been rapidly developed, and their packaging materials and technologies are well established.In this study, TGGT1_216200 (TG_200), a novel mol. from T. gondii, was identified using bioinformatic screening anal.TG_200 was purified and encapsulated with a lipid nanoparticle (LNP) to produce the TG_200 mRNA-LNP vaccine.The immune protection provided by the new vaccine and its mechanisms after immunizing BABL/C mice via i.m. injection were investigated.There was a strong immune response when mice were vaccinated with TG_200 mRNA-LNP.Elevated levels of anti-T. gondii-specific IgG (IgG), and a higher IgG2a-to-IgG1 ratio was observedThe levels of interleukin-12 (IL-12), interferon-γ (IFN-γ), IL-4, and IL-10 were also elevated.The result showed that the vaccine induced a mixture of Th1 and Th2 cells, and Th1-dominated humoral immune response.Significantly increased antigen-specific splenocyte proliferation was induced by TG_200 mRNA-LNP immunization.The vaccine could also induce T. gondii-specific cytotoxic T lymphocytes (CTLs).The expression levels of interferon regulatory factor 8 (IRF8), T-Box 21 (T-bet), and nuclear factor kappa B (NF-κB) were significantly elevated after TG_200 mRNA-LNP immunization.The levels of CD83, CD86, MHC-I, MHC-II, CD8, and CD4 mols. were also higher.The results indicated that TG_200 mRNA-LNP produced specific cellular and humoral immune responses.Most importantly, TG_200 mRNA-LNP immunized mice survived significantly longer (19.27 ± 3.438 days) than the control mice, which died within eight days after T. gondii challenge (P< 0.001).The protective effect of adoptive transfer was also assessed, and mice receiving serum and splenocytes from mice immunized with TG_200 mRNA-LNP showed improved survival rates of 9.70 ± 1.64 days and, 13.40 ± 2.32 days, resp. (P< 0.001).The results suggested that TG_200 mRNA-LNP is a safe and promising vaccine against T. gondii infection.

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Indication	Highest Phase	Country/Location	Organization	Date
Toxoplasmosis	Preclinical	China	Hangzhou Medical College	-

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TG_200 mRNA(Hangzhou Medical College)

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