Coléon Séverin, Assil Sonia, Dreux Marlène
CIRI, Inserm, U1111, Université Claude Bernard Lyon 1, CNRS, UMR5308, École Normale Supérieure de Lyon, Univ Lyon, Lyon, France.
Methods Mol Biol. 2019;1911:319-335. doi: 10.1007/978-1-4939-8976-8_22.
Plasmacytoid dendritic cells (pDCs) constitute a unique DC subset specialized in rapid and massive secretion of cytokines, including type I interferon (i.e., IFNα and IFNβ), known to be pivotal for both innate immunity and the onset of adaptive response. The production of type I IFNs by pDCs is primarily induced by the recognition of viral nucleic acids through Toll-like receptor (TLR)-7 and -9 sensors located in the endolysosomal compartment. Importantly, in the context of hepatitis C virus (HCV) infection, pDC type I IFN response is triggered by the sensing of infected cells via physical cell-cell contact. Such a feature is also observed for many genetically distant viruses, including notably viruses of the Retroviridae, Arenaviridae, Flaviviridae, Picornaviridaea, Togaviridae families and observed for various infected cell types. Here, we described a set of experimental methods for the ex vivo studies of the regulation of pDC activation upon physical cell-cell contact with virally infected cells.
浆细胞样树突状细胞(pDC)构成了一类独特的树突状细胞亚群,专门负责快速大量分泌细胞因子,包括I型干扰素(即IFNα和IFNβ),已知其对固有免疫和适应性反应的启动都至关重要。pDC产生I型干扰素主要是通过位于内溶酶体区室的Toll样受体(TLR)-7和-9传感器识别病毒核酸来诱导的。重要的是,在丙型肝炎病毒(HCV)感染的情况下,pDC的I型干扰素反应是通过物理性细胞间接触感知被感染细胞而触发的。许多亲缘关系较远的病毒也有这样的特征,包括逆转录病毒科、沙粒病毒科、黄病毒科、小RNA病毒科、披膜病毒科的病毒,并且在各种被感染的细胞类型中都观察到了这一现象。在此,我们描述了一套用于体外研究pDC与病毒感染细胞发生物理性细胞间接触时其激活调控的实验方法。