College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China; National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China.
Nanchong Key Laboratory of Disease Prevention, Control and Detection in Livestock and Poultry, Nanchong Vocational and Technical College, Nanchong 637131, China.
Res Vet Sci. 2019 Aug;125:382-389. doi: 10.1016/j.rvsc.2019.08.002. Epub 2019 Aug 5.
Toxoplasma gondii is an intracellular opportunistic, parasitic protozoan. Microglia have been classified into two main types: M1 (classically activated macrophages) and M2 (alternatively activated macrophages). BV2 cells were used in this study, together with lipopolysaccharide (LPS) and interferon (IFN)-γ or interleukin (IL)-4, which were used to induce resting microglia. Expression levels of M1/M2 markers were determined at both mRNA and protein levels, using PCR, western blot, and flow cytometry. Furthermore, cells were infected with T. gondii PLK strain, and the dynamic changes in M1/M2 marker expression levels were determined. An in vitro polarization model was successfully established. Expression of Nos2 and M1-associated markers was significantly upregulated at 12 h post-infection in BV2 cells. Further, the JAK/STAT1 and NF-κB signaling pathways were also activated following T. gondii infection. This demonstrated that T. gondii infection induces M1-type microglial polarization in vitro. The present study demonstrated that T. gondii infection affects microglial activation in vitro and elucidated the effects of activated microglia on T. gondii proliferation. This data may serve as a useful reference for more detailed elucidation of interactions between T. gondii and the innate immune system.
刚地弓形虫是一种细胞内机会性、寄生性的原生动物。小胶质细胞分为两种主要类型：M1（经典激活的巨噬细胞）和 M2（交替激活的巨噬细胞）。本研究使用 BV2 细胞，并用脂多糖（LPS）和干扰素（IFN）-γ或白细胞介素（IL）-4 诱导静止小胶质细胞。使用 PCR、western blot 和流式细胞术在 mRNA 和蛋白质水平上测定 M1/M2 标志物的表达水平。此外，用刚地弓形虫 PLK 株感染细胞，并测定 M1/M2 标志物表达水平的动态变化。成功建立了体外极化模型。在感染后 12 小时，BV2 细胞中 Nos2 和 M1 相关标志物的表达显著上调。此外，刚地弓形虫感染还激活了 JAK/STAT1 和 NF-κB 信号通路。这表明刚地弓形虫感染可诱导体外 M1 型小胶质细胞极化。本研究表明，刚地弓形虫感染可影响体外小胶质细胞的激活，并阐明了激活的小胶质细胞对刚地弓形虫增殖的影响。这些数据可能为更详细地阐明刚地弓形虫与先天免疫系统之间的相互作用提供有用的参考。
