Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Department of Parasitology and Mycology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
Immun Inflamm Dis. 2024 Jun;12(6):e1329. doi: 10.1002/iid3.1329.
Toxoplasma gondii is an obligate intracellular protozoan parasite that can invade all mammalian cells. It is well established that natural killer (NK) cells have critical protective roles in innate immunity during infections by intracellular pathogens. In the current study, we conducted an in vitro experiment to evaluate NK cell differentiation and activation from human umbilical cord blood mononuclear cells (UCB-MNCs) after infection with T. gondii tachyzoites.
UCB-MNCs were infected by fresh tachyzoites of type I (RH) or type II (PTG) strains of T. gondii pre-expanded in mesenchymal stem cells for 2 weeks in a medium enriched with stem cell factor, Flt3, IL-2, and IL-15. Flow cytometry analysis and western blot analysis were performed to measure the CD57, CD56, and Granzyme A (GZMA).
Data revealed that incubation of UCB-MNCs with NK cell differentiation medium increased the CD57, CD56, and GZMA. UCB-MNCs cocultured with PTG tachyzoites showed a significant reduction of CD56 and GZMA, but nonsignificant changes, in the levels of CD56 compared to the control UCB-MNCs (p > .05). Noteworthy, 2-week culture of UCB-MNCs with type I (RH) tachyzoites significantly suppressed CD57, CD56, and GZMA, showing reduction of NK cell differentiation from cord blood cells.
Our findings suggest that virulent T. gondii tachyzoites with cytopathic effects inhibit NK cell activation and eliminate innate immune responses during infection, and consequently enable the parasite to continue its survival in the host body.
刚地弓形虫是一种专性细胞内原生动物寄生虫,能够侵入所有哺乳动物细胞。众所周知,自然杀伤 (NK) 细胞在细胞内病原体感染期间的固有免疫中具有关键的保护作用。在目前的研究中,我们进行了一项体外实验,以评估感染刚地弓形虫速殖子后,从人脐血单核细胞 (UCB-MNC) 中分化和激活 NK 细胞。
UCB-MNC 用新鲜的 I 型 (RH) 或 II 型 (PTG) 刚地弓形虫速殖子感染,这些速殖子在富含干细胞因子、Flt3、IL-2 和 IL-15 的培养基中预先在间充质干细胞中扩增 2 周。通过流式细胞术分析和 Western blot 分析测量 CD57、CD56 和颗粒酶 A (GZMA)。
数据显示,用 NK 细胞分化培养基孵育 UCB-MNC 会增加 CD57、CD56 和 GZMA。与对照 UCB-MNC 相比,与 PTG 速殖体共培养的 UCB-MNC 显示 CD56 和 GZMA 显著减少,但 CD56 水平没有显著变化 (p>.05)。值得注意的是,用 I 型 (RH) 速殖体培养 2 周的 UCB-MNC 显著抑制 CD57、CD56 和 GZMA,表明从脐血细胞中分化 NK 细胞减少。
我们的研究结果表明,具有细胞病变效应的毒力刚地弓形虫速殖子抑制 NK 细胞激活并消除感染期间的固有免疫反应,从而使寄生虫能够继续在宿主体内生存。