Student Research Committee, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Department of Medical Parasitology and Mycology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Foodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Microb Pathog. 2021 Sep;158:105072. doi: 10.1016/j.micpath.2021.105072. Epub 2021 Jun 27.
Toxoplasma gondii (T. gondii) is an intracellular parasitic protozoan infecting homoeothermic animals and about a third of the world's population. Inflammasomes are intracellular multi-protein complex, which are activated by many factors. Inflammasomes are activated during toxoplasmosis; however, there are a lot of obscure aspects. THP-1 monocyte cells were converted to M0 macrophages by PMA and treated by 100 μg/mL soluble total Ag (STAg) derived from T. gondii strain RH for two time points 3 h and 24 h. After total RNA extraction and cDNA synthesis, the expression pattern of NLRP1, NLRP3, NLRC4, AIM2, IL1β, and IL18 was evaluated by relative real-time PCR. In addition, the cytokine release of IL1β and TNFα was evaluated in the supernatant of each well. The results showed statistically significant time-dependent overexpression of inflammasomes. NLRP1 and NLRP3 showed the higher and lower expression, respectively, during 3 h and 24 h after exposure. Both IL1β and IL18 downregulated 3 h after exposure. IL18 presented statistically significant upregulation after 24 h, but IL1β showed statistically significant downregulation after 24 h. The release of IL1β increased after 3 h, but it slightly decreased during 24 h after exposure. The concentration of TNFα showed an insignificant decrease compared to control, while it increased during 24 h after exposure. Taken together, this study suggested that T. gondii STAg induces NLRP1 more than NLRP3, NLRC4, and AIM2. Our findings also proposed that T. gondii STAg downregulates the gene expression of IL1β, but increases the release of this cytokine. It seems that Toxoplasma STAg probably increase the release of IL1β via activating NLRPs and AIM2 to cleave pro-caspase 1 to caspase 1 that leads to conversion of pro IL1β to mature IL1β.
刚地弓形虫(Toxoplasma gondii)是一种感染恒温动物和世界上约三分之一人口的细胞内寄生原生动物。炎症小体是一种细胞内多蛋白复合物,可被多种因素激活。在弓形虫感染期间激活炎症小体; 然而,还有很多方面尚不清楚。通过 PMA 将 THP-1 单核细胞转化为 M0 巨噬细胞,并分别用 100μg/mL 的 T. gondii RH 株可溶性总 Ag(STAg)处理 3h 和 24h。提取总 RNA 并合成 cDNA 后,通过相对实时 PCR 评估 NLRP1、NLRP3、NLRC4、AIM2、IL1β 和 IL18 的表达模式。此外,还评估了每个孔上清液中细胞因子 IL1β 和 TNFα 的释放。结果显示,炎症小体的表达呈现出统计学上显著的时间依赖性上调。NLRP1 和 NLRP3 在暴露后 3h 和 24h 时分别表现出更高和更低的表达。IL1β 和 IL18 在暴露后 3h 下调。IL18 在暴露后 24h 时呈现统计学显著上调,但 IL1β 在暴露后 24h 时呈现统计学显著下调。IL1β 的释放量在 3h 后增加,但在暴露后 24h 时略有下降。与对照相比,TNFα 的浓度呈下降趋势,但在暴露后 24h 时增加。综上所述,本研究表明 T. gondii STAg 诱导 NLRP1 的表达高于 NLRP3、NLRC4 和 AIM2。我们的研究结果还表明,T. gondii STAg 下调了 IL1β 的基因表达,但增加了这种细胞因子的释放。似乎 Toxoplasma STAg 可能通过激活 NLRPs 和 AIM2 增加 IL1β 的释放,从而将前体 caspase 1 切割为 caspase 1,导致前体 IL1β 转化为成熟的 IL1β。