Department of Mycotic and Parasitic Agents and Mycobacteria, Robert Koch-Institut, Berlin, Germany.
Center for Biological Threats and Special Pathogens, Highly Pathogenic Viruses, Robert Koch-Institut, Berlin, Germany.
Sci Rep. 2018 Feb 12;8(1):2797. doi: 10.1038/s41598-018-21143-0.
Rodent species like Myodes glareolus and Microtus spp. are natural reservoirs for many zoonotic pathogens causing human diseases and are gaining increasing interest in the field of eco-immunology as candidate animal models. Despite their importance the lack of immunological reagents has hampered research in these animal species. Here we report the recombinant production and functional characterization of IFN-γ, a central mediator of host's innate and adaptive immune responses, from the bank vole M. glareolus. Soluble dimeric recMgIFN-γ was purified in high yield from Escherichia coli. Its activity on M. glareolus and Microtus arvalis kidney cell lines was assessed by immunofluorescent detection of nuclear translocation and phosphorylation of the transcription factor STAT1. RecMgIFN-γ also induced expression of an IFN-γ-regulated innate immunity gene. Inhibition of vesicular stomatitis virus replication in vole cells upon recMgIFN-γ treatment provided further evidence of its biological activity. Finally, we established a recMgIFN-γ-responsive bank vole reporter cell line that allows the sensitive titration of the cytokine activity via a bioluminescence reporter assay. Taken together, we report valuable tools for future investigations on the immune response against zoonotic pathogens in their natural animal hosts, which might foster the development of novel animal models.
像黑线姬鼠和田鼠属的啮齿动物物种是许多引起人类疾病的人畜共患病原体的天然宿主，它们作为候选动物模型在生态免疫学领域引起了越来越多的关注。尽管它们很重要，但缺乏免疫试剂阻碍了这些动物物种的研究。在这里，我们报告了来自黑线姬鼠 M. glareolus 的 IFN-γ的重组生产和功能表征，IFN-γ 是宿主固有和适应性免疫反应的中心介质。从大肠杆菌中以高产量纯化了可溶性二聚体 recMgIFN-γ。通过免疫荧光检测转录因子 STAT1 的核易位和磷酸化来评估 recMgIFN-γ 对黑线姬鼠和黑线仓鼠肾细胞系的活性。RecMgIFN-γ 还诱导了 IFN-γ 调节的先天免疫基因的表达。在 recMgIFN-γ 处理后，抑制水疱性口炎病毒在田鼠细胞中的复制提供了其生物学活性的进一步证据。最后，我们建立了一种 recMgIFN-γ 反应性的黑线姬鼠报告细胞系，该细胞系允许通过生物发光报告测定法灵敏地滴定细胞因子活性。总之，我们报告了针对其天然动物宿主中人畜共患病原体的免疫反应的有价值的工具，这可能会促进新型动物模型的开发。
