Zhang Zhi-Wei, Li Ting-Ting, Wang Jin-Lei, Liang Qin-Li, Zhang Hai-Sheng, Sun Li-Xiu, Zhu Xing-Quan
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.
College of Veterinary Medicine, Shanxi Agricultural University, Taigu, China.
Front Vet Sci. 2021 Jan 14;7:614759. doi: 10.3389/fvets.2020.614759. eCollection 2020.
Toxoplasmosis caused by infection with is an important parasitic zoonosis with a worldwide distribution. In this study, we examined the functions of two thioredoxins (namely CTrp26 and CTrx1) of tachyzoites by generation of HA tag strains or gene deficient parasites in Type I RH strain (ToxoDB#10). Immunofluorescence analysis (IFA) was used to investigate the subcellular localization of the thioredoxins (Trxs). Results of IFA showed that both CTrp26 and CTrx1 were located in the cytoplasm of . Functional characterizations of CTrp26 and CTrx1-deficient parasites were performed by plaque assay, intracellular replication, egress, HO resistance, detection of reactive oxygen species (ROS) level and total antioxidant capacity (T-AOC) assays , as well as mouse infection . Our results showed that deletion of CTrp26 or CTrx1 did not influence the ability of RH strain to replicate, egress, form plaque, resist HO exposure, maintain the ROS level, and T-AOC, and also did not serve as virulence factors in Kunming mice. Taken together, these results provide new properties of the two Trxs. Although they are not essential for RH strain, they may have roles in other strains of this parasite due to their different expression patterns, which warrants future research.
由 感染引起的弓形虫病是一种重要的寄生虫人畜共患病，在全球范围内均有分布。在本研究中，我们通过在I型RH株（ToxoDB#10）中构建HA标签菌株或基因缺陷型寄生虫，研究了速殖子的两种硫氧还蛋白（即CTrp26和CTrx1）的功能。采用免疫荧光分析（IFA）研究硫氧还蛋白（Trxs）的亚细胞定位。IFA结果显示，CTrp26和CTrx1均位于 的细胞质中。通过噬斑试验、细胞内复制、逸出、对HO的抗性、活性氧（ROS）水平检测、总抗氧化能力（T-AOC）测定以及小鼠感染实验，对CTrp26和CTrx1缺陷型寄生虫进行功能表征。我们的结果表明，缺失CTrp26或CTrx1并不影响RH株的复制、逸出、形成噬斑、抵抗HO暴露、维持ROS水平和T-AOC的能力，在昆明小鼠中也不作为毒力因子。综上所述，这些结果揭示了这两种硫氧还蛋白的新特性。尽管它们对RH株并非必需，但由于其不同的表达模式，可能在该寄生虫的其他菌株中发挥作用，值得未来进一步研究。
