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

Ma, Zhen; Yan, Kang; Jiang, Ruo-Lan; Guan, Jie; Yang, Le; Huang, Yehong; Lü, Bin; Li, Xuanwu; Zhang, Jie; Chang, Yunfeng; Wu, Xiang

doi:10.3389/fmicb.2020.00399

Cited by 4 publications

(5 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Based on our previous studies, we found that the wx2 gene is associated with the virulence of T. gondii strains and may be involved in regulating the host immune response [ 32 ]. In this study, the virulence assay of mice revealed that the survival time for mice infected with wx2 gene knockout strain RH wx2−/− was significantly longer than that for the mice infected with the wild-type RH strain.…”

Section: Discussionmentioning

confidence: 99%

“…Tachyzoites of T. gondii RH strain (type I) and wx2 knockout (RH wx2−/− strain) were stored in our laboratory. The wx2 gene complementary strains were constructed as described in the literature using CRISPR-Cas9 [ 32 , 33 ]. Ptub::GOI::CAT plasmid with the wx2 fragment and purified wx2 -CAT fragment were co-transfected into freshly harvested tachyzoites of RH wx2−/− strain by electroporation.…”

Section: Methodsmentioning

confidence: 99%

“…Our previous studies have shown that the wx2 gene may be related to T. gondii virulence and may even regulate the host immune response to achieve immune escaping [ 32 ]. In this study, we explore whether the wx2 gene of T. gondii affects the host immune response via the pyroptosis pathway in vitro and in vivo.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Virulence-related gene wx2 of Toxoplasma gondii regulated host immune response via classic pyroptosis pathway

Jiang

et al. 2022

Parasites Vectors

Self Cite

View full text Add to dashboard Cite

Background 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. Methods The wx2 knockout strain (RHwx2−/− strain) and complementary strain (RHwx2+/+ 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. Results Compared with the wild RH and RHwx2+/+ strain groups, the survival time for mice infected with the RHwx2−/− 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 RHwx2−/− strain increased to different degrees, compared with infected with wild RH strain and RHwx2+/+ 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 RHwx2−/− were significantly higher than those of RH and RHwx2+/+ strains, suggesting that the wx2 may inhibit the host's immune response. Conclusion wx2 is a virulence related gene of T. gondii, and may be involved in host immune regulation by inhibiting the pyroptosis pathway. Graphical Abstract

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Virulence-related gene wx2 of Toxoplasma gondii regulated host immune response via classic pyroptosis pathway

Jiang

et al. 2022

Parasites Vectors

Self Cite

View full text Add to dashboard Cite

show abstract

“…Th17 cells play dual functions in the process of T. gondii infection. In the early stage of infection, Th17 can resist the invasion and damage of T. gondii , while in the later stage, Th17 can trigger immune pathological changes in the body, resulting in severe inflammatory response ( Ma et al., 2020 ). Clinical studies have found that Th17 cells and the proinflammatory factor IL-17 are increased in toxoplasmosis ( Dutra et al., 2013 ; Silva et al., 2014 ), and IL-17 will further promote the inflammatory response and expand tissue damage after T. gondii infection ( Guiton et al., 2010 ).…”

Section: Th17 Cells and Mood Disordersmentioning

confidence: 99%

From the immune system to mood disorders especially induced by Toxoplasma gondii: CD4+ T cell as a bridge

Wang

Zhong

Chen

et al. 2023

Front. Cell. Infect. Microbiol.

View full text Add to dashboard Cite

Toxoplasma gondii (T. gondii), a ubiquitous and obligatory intracellular protozoa, not only alters peripheral immune status, but crosses the blood-brain barrier to trigger brain parenchymal injury and central neuroinflammation to establish latent cerebral infection in humans and other vertebrates. Recent findings underscore the strong correlation between alterations in the peripheral and central immune environment and mood disorders. Th17 and Th1 cells are important pro-inflammatory cells that can drive the pathology of mood disorders by promoting neuroinflammation. As opposed to Th17 and Th1, regulatory T cells have inhibitory inflammatory and neuroprotective functions that can ameliorate mood disorders. T. gondii induces neuroinflammation, which can be mediated by CD4+ T cells (such as Tregs, Th17, Th1, and Th2). Though the pathophysiology and treatment of mood disorder have been currently studied, emerging evidence points to unique role of CD4+ T cells in mood disorder, especially those caused by T. gondii infection. In this review, we explore some recent studies that extend our understanding of the relationship between mood disorders and T. gondii.

show abstract

“…For positive selection of T. gondii mutant or transgenic strains that have successfully integrated an exogenous DNA coding for a modified dihydrofolate reductase-thymidylate synthase (mDHFR-TS), pyrimethamine (Pyr) is the drug of choice [15][16][17][18][19], since mDHFR-TS confers resistance to Pyr [20]. In the genome of WT T. gondii, a single-copy gene coding for DHFR-TS (WT-DHFR-TS) is expressed, but the enzyme is sensitive to Pyr [21].…”

Section: Introductionmentioning

confidence: 99%

Single- and duplex TaqMan-quantitative PCR for determining the copy numbers of integrated selection markers during site-specific mutagenesis in Toxoplasma gondii by CRISPR-Cas9

et al. 2022

View full text Add to dashboard Cite

Herein, we developed a single and a duplex TaqMan quantitative PCR (qPCR) for absolute quantification of copy numbers of integrated dihydrofolate reductase-thymidylate synthase (mdhfr-ts) drug selectable marker for pyrimethamine resistance in Toxoplasma gondii knockouts (KOs). The single TaqMan qPCR amplifies a 174 bp DNA fragment of the inserted mdhfrts and of the wild-type (WT) dhfr-ts (wtdhfr-ts) which is present as single copy gene in Toxoplasma and encodes a sensitive enzyme to pyrimethamine. Thus, the copy number of the dhfr-ts fragment in a given DNA quantity from KO parasites with a single site-specific integration should be twice the number of dhfr-ts copies recorded in the same DNA quantity from WT parasites. The duplex TaqMan qPCR allows simultaneous amplification of the 174 bp dhfr-ts fragment and the T. gondii 529-bp repeat element. Accordingly, for a WT DNA sample, the determined number of tachyzoites given by dhfr-ts amplification is equal to the number of tachyzoites determined by amplification of the Toxoplasma 529-bp, resulting thus in a ratio of 1. However, for a KO clone having a single site-specific integration of mdhfr-ts, the calculated ratio is 2. We then applied both approaches to test T. gondii RH mutants in which the major surface antigen (SAG1) was disrupted through insertion of mdhfr-ts using CRISPR-Cas9. Results from both assays were in correlation showing a high accuracy in detecting KOs with multiple integrated mdhfr-ts. Southern blot analyses using BsaBI and DraIII confirmed qPCRs results. Both TaqMan qPCRs are needed for reliable diagnostic of T. gondii KOs following CRISPR-Cas9-mediated mutagenesis, particularly with respect to off-target effects resulting from multiple insertions of mdhfr-ts. The principle of the duplex TaqMan qPCR is applicable for other selectable markers in Toxoplasma. TaqMan qPCR tools may contribute to more frequent use of WT Toxoplasma strains during functional genomics.

show abstract

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

Abstract: (2020) A Novel wx2 Gene of Toxoplasma gondii Inhibits the Parasitic Invasion and Proliferation in vitro and Attenuates Virulence in vivo via Immune Response Modulation.

Cited by 4 publications

References 32 publications

Virulence-related gene wx2 of Toxoplasma gondii regulated host immune response via classic pyroptosis pathway

Virulence-related gene wx2 of Toxoplasma gondii regulated host immune response via classic pyroptosis pathway

From the immune system to mood disorders especially induced by Toxoplasma gondii: CD4+ T cell as a bridge

Single- and duplex TaqMan-quantitative PCR for determining the copy numbers of integrated selection markers during site-specific mutagenesis in Toxoplasma gondii by CRISPR-Cas9

Contact Info

Product

Resources

About