biostudies-literatureMa ZNational Natural Sciences Foundation of China454https://www.ebi.ac.uk/biostudies/studies/S-EPMC9724370biostudies-literatureUnknown15(1)<h4>Background</h4>Toxoplasma gondii is known as the most successful parasite, which can regulate the host immune response through a variety of ways to achieve immune escape. We previously reported that a novel gene wx2 of T. gondii may be a virulence-related molecule. The objective of this study was to explore the mechanism of wx2 regulating host immune response.<h4>Methods</h4>The wx2 knockout strain (RH^wx2-/- strain) and complementary strain (RH^wx2+/+ strain) were constructed by the CRISPR/Cas9 technique, and the virulence of the wx2 gene was detected and changes in pyroptosis-related molecules were observed.<h4>Results</h4>Compared with the wild RH and RH^wx2+/+ strain groups, the survival time for mice infected with the RH^wx2-/- strain was prolonged to a certain extent. The mRNA levels of pyroptosis-related molecules of caspase-1, NLRP3, and GSDMD and et al. in mouse lymphocytes in vivo and RAW267.4 cells in vitro infected with RH^wx2-/- strain increased to different degrees, compared with infected with wild RH strain and RH^wx2+/+ strain. As with the mRNA level, the protein level of caspase-1, caspase-1 p20, IL-1β, NLRP3, GSDMD-FL, GSDMD-N, and phosphorylation level of NF-κB (p65) were also significantly increased. These data suggest that wx2 may regulate the host immune response through the pyroptosis pathway. In infected RAW264.7 cells at 48 h post-infection, the levels of Th1-type cytokines of IFN-γ, Th2-type cytokines such as IL-13, Th17-type cytokine of IL-17 in cells infected with RH^wx2-/- were significantly higher than those of RH and RH^wx2+/+ strains, suggesting that the wx2 may inhibit the host's immune response.<h4>Conclusion</h4>wx2 is a virulence related gene of T. gondii, and may be involved in host immune regulation by inhibiting the pyroptosis pathway.Parasites & vectorsVirulence-related gene wx2 of Toxoplasma gondii regulated host immune response via classic pyroptosis pathway.PMC972437082072306Li XYan KLi ZHuang YJiang RZhang NMa ZLu BDibo NWu XfalseVirulence-related gene wx2 of Toxoplasma gondii regulated host immune response via classic pyroptosis pathway.<h4>Background</h4>Toxoplasma gondii is known as the most successful parasite, which can regulate the host immune response through a variety of ways to achieve immune escape. We previously reported that a novel gene wx2 of T. gondii may be a virulence-related molecule. The objective of this study was to explore the mechanism of wx2 regulating host immune response.<h4>Methods</h4>The wx2 knockout strain (RH^wx2-/- strain) and complementary strain (RH^wx2+/+ strain) were constructed by the CRISPR/Cas9 technique, and the virulence of the wx2 gene was detected and changes in pyroptosis-related molecules were observed.<h4>Results</h4>Compared with the wild RH and RH^wx2+/+ strain groups, the survival time for mice infected with the RH^wx2-/- strain was prolonged to a certain extent. The mRNA levels of pyroptosis-related molecules of caspase-1, NLRP3, and GSDMD and et al. in mouse lymphocytes in vivo and RAW267.4 cells in vitro infected with RH^wx2-/- strain increased to different degrees, compared with infected with wild RH strain and RH^wx2+/+ strain. As with the mRNA level, the protein level of caspase-1, caspase-1 p20, IL-1β, NLRP3, GSDMD-FL, GSDMD-N, and phosphorylation level of NF-κB (p65) were also significantly increased. These data suggest that wx2 may regulate the host immune response through the pyroptosis pathway. In infected RAW264.7 cells at 48 h post-infection, the levels of Th1-type cytokines of IFN-γ, Th2-type cytokines such as IL-13, Th17-type cytokine of IL-17 in cells infected with RH^wx2-/- were significantly higher than those of RH and RH^wx2+/+ strains, suggesting that the wx2 may inhibit the host's immune response.<h4>Conclusion</h4>wx2 is a virulence related gene of T. gondii, and may be involved in host immune regulation by inhibiting the pyroptosis pathway.2022-01-01T00:00:00Z2022 Dec2025-04-19T13:22:09.833Z2025-04-19T13:22:09.833ZS-EPMC97243703647141710.1186/s13071-022-05502-5