rhoptry 蛋白 9 的缺失阻碍了刚地弓形虫的感染性。
Loss of rhoptry protein 9 impeded Toxoplasma gondii infectivity.
机构信息
Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, China; Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry, College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin 300384, China.
Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, China; The Research Unit for Pathogenic Mechanisms of Zoonotic Parasites, Chinese Academy of Medical Sciences, 120 Dongling Road, Shenyang 110866, China.
出版信息
Acta Trop. 2020 Jul;207:105464. doi: 10.1016/j.actatropica.2020.105464. Epub 2020 Apr 14.
Toxoplasma gondii is an obligatory intracellular parasite that critically depends on active invasion and egress from infected host cells to complete its propagation cycle. T. gondii rhoptry proteins (TgROPs) are virulent factors associated with host cell invasion, growth. In this study, we analyzed the functions of ROP9 in the process of T. gondii infection. The TgROP9 knockout RH strain (RH△ROP9) and its recovery strain (RH-ReROP9) were constructed using the CRISPR/Cas9 system. The invasion, proliferation, and egress efficiency of the RH△ROP9 strain were evaluated and their pathogenicity to mice was analyzed. Compared with RH wild-type (RH-WT) and RH-ReROP9 strains, the invasion percentage of RH△ROP9 to Vero cells was reduced by about 28.0% (p< 0.01) at 1.5 h, and the relative proliferation percentage was decreased by about 35.0% (p< 0.01) after infection with 10 or 10 parasites. In addition, the RH△ROP9 strain also showed prolonged egress time from host cells. The survival time of the mice (12.6 ± 1.6 or 10.1 ± 1.1 days) were delayed (p < 0.001) after infection with either 200 or 1000 RH△ROP9 parasites. These evidences suggested that ROP9 facilitated T. gondii infection in vitro and in vivo.
刚地弓形虫是一种必需的细胞内寄生虫，严重依赖于从感染宿主细胞中的主动入侵和逸出，以完成其繁殖周期。刚地弓形虫的 rhoptry 蛋白（TgROPs）是与宿主细胞入侵、生长相关的毒力因子。在本研究中，我们分析了 ROP9 在刚地弓形虫感染过程中的功能。使用 CRISPR/Cas9 系统构建了 TgROP9 敲除 RH 株（RH△ROP9）及其恢复株（RH-ReROP9）。评估了 RH△ROP9 株的入侵、增殖和逸出效率，并分析了其对小鼠的致病性。与 RH 野生型（RH-WT）和 RH-ReROP9 株相比，RH△ROP9 株对 Vero 细胞的入侵率在 1.5 h 时降低了约 28.0%（p<0.01），感染 10 或 10 个寄生虫后相对增殖率降低了约 35.0%（p<0.01）。此外，RH△ROP9 株还表现出从宿主细胞中延长的逸出时间。感染 200 或 1000 RH△ROP9 寄生虫后，小鼠的存活时间（12.6±1.6 或 10.1±1.1 天）延长（p<0.001）。这些证据表明 ROP9 促进了刚地弓形虫在体外和体内的感染。
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