Bruchhagen Christin, van Krüchten Andre, Klemm Carolin, Ludwig Stephan, Ehrhardt Christina
Institute of Virology Muenster (IVM), Westfaelische Wilhelms-University Muenster, Muenster, Germany.
Methods Mol Biol. 2018;1836:375-386. doi: 10.1007/978-1-4939-8678-1_18.
Investigation of pathogen-host interactions on a molecular level requires sophisticated in vitro infection procedures, especially in the presence of different pathogens.Super-infections of influenza viruses (IV) and bacteria, with increasing incidence of Staphylococcus aureus (S. aureus) cases, are a long-known phenomenon and represent a major complication in IV-infected patients. Although several in vivo studies have improved our knowledge about pathogenesis and immune responses of super-infections that result in increased morbidity and mortality, the consequences of the direct interplay of viruses and bacteria on a molecular level in affected cells that may contribute to the deadly synergism of these pathogens are so far poorly characterized. Here we describe different infection schemes to study IV and S. aureus coinfections of distinct cell populations in vitro. Depending on the focus of interest, regulation of cell responses such as signalling mechanisms or pro- and anti-inflammatory cytokine expression, or consequences for the viral or bacterial life cycle, can be analyzed. The described infection procedures could be used as guidelines and adapted to super-infection settings of other viral and bacterial pathogens.
在分子水平上研究病原体与宿主的相互作用需要复杂的体外感染程序,尤其是在存在不同病原体的情况下。流感病毒(IV)与细菌的重叠感染,随着金黄色葡萄球菌(S. aureus)病例的增加,是一个早已为人所知的现象,并且是IV感染患者的主要并发症。尽管一些体内研究增进了我们对导致发病率和死亡率增加的重叠感染的发病机制和免疫反应的了解,但病毒和细菌在受影响细胞中分子水平上的直接相互作用的后果,可能导致这些病原体致命协同作用,到目前为止其特征还不清楚。在这里,我们描述了不同的感染方案,以研究IV和金黄色葡萄球菌在体外对不同细胞群体的共感染。根据研究重点,可以分析细胞反应的调节,如信号传导机制或促炎和抗炎细胞因子的表达,或对病毒或细菌生命周期的影响。所描述的感染程序可作为指导方针,并适用于其他病毒和细菌病原体的重叠感染情况。