Oltean Teodora, Maelfait Jonathan, Saelens Xavier, Vandenabeele Peter
VIB Center for Inflammation Research, Cell Death and Inflammation Unit, Ghent, Belgium.
Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
Cell Death Discov. 2024 May 22;10(1):247. doi: 10.1038/s41420-024-01981-w.
The involvement of necroptosis in the control of influenza A virus (IAV) infection has been reported in multiple studies. Downstream of the nucleic acid sensor ZBP1, RIPK3 kinase activity is critically involved in the induction of necroptotic cell death by phosphorylating MLKL, while RIPK3 as a scaffold can induce apoptosis. Paradoxically, RIPK3-deficiency of mice may result in increased or decreased susceptibility to IAV infection. Here, we critically review the published reports on the involvement of RIPK3 in IAV infection susceptibility and try to identify differences in experimental settings that could explain seemingly conflicting outcomes. Analysis of the experimental reports revealed differences in the IAV challenge dose, the IAV inoculum preparation, IAV titer assessment, as well as the route of inoculation between studies. Furthermore, differences were noticed in the inclusion of littermate controls, which show high variance in viral sensitivity. Our evaluation argues for a standardized setup for IAV infection experiments including the preparation of the IAV virus, the use of different IAV infectious doses description and the proper experimental genetic controls of the mouse strains to increase inter-laboratory consistency in this field. Workflow for IAV infection studies in vivo: Viral preparation and titer assessment should be as standardized as possible with the use of a universal repository (such as BEI resources). Infection studies in genetically modified mice and littermate controls should include dose-response experimentation, following a defined infection route and inoculation volume. Data are generated by consistent analysis methods.
多项研究报道了坏死性凋亡在甲型流感病毒（IAV）感染控制中的作用。在核酸传感器ZBP1的下游，RIPK3激酶活性通过磷酸化MLKL在坏死性凋亡细胞死亡的诱导中起关键作用，而RIPK3作为一种支架蛋白可诱导凋亡。矛盾的是，RIPK3基因敲除小鼠对IAV感染的易感性可能增加或降低。在此，我们严格审查了已发表的关于RIPK3在IAV感染易感性中作用的报告，并试图找出实验设置中的差异，这些差异可以解释看似相互矛盾的结果。对实验报告的分析揭示了不同研究之间在IAV攻毒剂量、IAV接种体制备、IAV滴度评估以及接种途径方面存在差异。此外，在同窝对照的纳入方面也存在差异，同窝对照显示出病毒敏感性的高度变异性。我们的评估主张对IAV感染实验进行标准化设置，包括IAV病毒的制备、不同IAV感染剂量的描述以及小鼠品系适当的实验遗传对照，以提高该领域实验室间的一致性。体内IAV感染研究的工作流程：病毒制备和滴度评估应尽可能标准化，使用通用储存库（如BEI资源）。在基因改造小鼠和同窝对照中的感染研究应包括剂量反应实验，遵循确定的感染途径和接种体积。数据通过一致的分析方法生成。
