Yang Shuo, Zhao Yaya, Yu Junjing, Fan Zhiqin, Gong Si-Tang, Tang Hong, Pan Lei
The Joint Center for Infection and Immunity, Guangzhou Institute of Pediatrics, Guangzhou Women and Children's Medical Center; CAS Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences; CAS Key Laboratory of Infection and Immunity (CASKLII), Institute of Biophysics, Chinese Academy of Sciences.
The Joint Center for Infection and Immunity, Guangzhou Institute of Pediatrics, Guangzhou Women and Children's Medical Center; CAS Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences.
J Vis Exp. 2019 Mar 14(145). doi: 10.3791/58845.
Virus spreading is a major cause of epidemic diseases. Thus, understanding the interaction between the virus and the host is very important to extend our knowledge of prevention and treatment of viral infection. The fruit fly Drosophila melanogaster has proven to be one of the most efficient and productive model organisms to screen for antiviral factors and investigate virus-host interaction, due to powerful genetic tools and highly conserved innate immune signaling pathways. The procedure described here demonstrates a nano-injection method to establish viral infection and induce systemic antiviral responses in adult flies. The precise control of the viral injection dose in this method enables high experimental reproducibility. Protocols described in this study include the preparation of flies and the virus, the injection method, survival rate analysis, the virus load measurement, and an antiviral pathway assessment. The influence effects of viral infection by the flies' background were mentioned here. This infection method is easy to perform and quantitatively repeatable; it can be applied to screen for host/viral factors involved in virus-host interaction and to dissect the crosstalk between innate immune signaling and other biological pathways in response to viral infection.
病毒传播是传染病的主要原因。因此，了解病毒与宿主之间的相互作用对于扩展我们对病毒感染预防和治疗的认识非常重要。由于强大的遗传工具和高度保守的先天免疫信号通路，果蝇已被证明是筛选抗病毒因子和研究病毒-宿主相互作用最有效和高产的模式生物之一。本文所述的方法展示了一种通过纳米注射在成年果蝇中建立病毒感染并诱导全身抗病毒反应的方法。该方法中对病毒注射剂量的精确控制可实现高实验重复性。本研究中描述的方案包括果蝇和病毒的制备、注射方法、存活率分析、病毒载量测量以及抗病毒途径评估。这里提到了果蝇背景对病毒感染的影响。这种感染方法易于操作且可定量重复；它可用于筛选参与病毒-宿主相互作用的宿主/病毒因子，并剖析先天免疫信号与其他生物途径之间在病毒感染反应中的相互作用。
