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Toxoplasma gondii  ( T. gondii ) is an obligate intracellular apicomplexan protozoan that can parasitize most warm-blooded animals and cause severe diseases in immunocompromised individuals or fetal abnormalities in pregnant woman. The treatment of toxoplamosis has been limited by effective drugs. Our previous work indicated that the novel gene  wx2  of  T. gondii  may serve as a vaccine antigen candidate. To further investigate the molecular functions of  wx2  in highly virulent  T. gondii  (RH strain), a  wx2  gene deletion mutant RH strain (KO- wx2 ) was established using CRISPR-Cas9. The phynotype of KO- wx2  was analyzed by plaque, invasion, and replication assays  in vitro  as well as  in vivo  virulence assays. The results indicated that the targeted deletion of the  wx2  gene significantly inhibited  in vitro  parasite growth and replication in the host cells as well as attenuated parasite virulence in the mouse model. Notably, the percentage of pro-inflammatory factors of interferon gamma (IFN-γ) and interlukin-17A (IL-17A) and anti-inflammatory factor of interlukin-10 (IL-10) in the lymph nodes were upregulated in mice infected with the KO- wx2  strain. Our data suggested that the  wx2  gene plays an important role in the process of the parasite’s life cycle and virulence in mice. In addition, it also plays an important role in the host’s immunity reaction, mainly via Th1 and Th17 cellular immunity, not Th2.

frontiers in microbiology

A Novel wx2 Gene of Toxoplasma gondii Inhibits the Parasitic Invasion and Proliferation in vitro and Attenuates Virulence in vivo via Immune Response Modulation