Detecting single cell interferon-beta production using a fluorescent reporter telomerase-immortalized human fibroblast cell line

• Published in: STAR protocols Vol. 2; no. 2; p. 100436
• Main Authors: Hare, David N., Subapanditha, Minomi K., Mossman, Karen L.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 18.06.2021; Elsevier
• Subjects: Cell Biology; Cell-based Assays; Flow Cytometry/Mass Cytometry; Immunology; Microscopy; Protocol; Single Cell; Single Cell; Immunology; Microscopy; Flow Cytometry/Mass Cytometry; Cell-based Assays; Cell Biology
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2021.100436

Recent data suggest that cells respond to infection by upregulating the antiviral cytokine interferon-beta (IFN-ß) in a fraction of infected cells. Approaches are thus needed to study these responses on a single-cell level rather than bulk population. Here, we describe a protocol to analyze the IFN-ß response of individual cells using flow cytometry and immunofluorescence microscopy. We show the heterogeneous IFN-ß response to inactivated Sendai virus and human cytomegalovirus, but this protocol can be adapted to other viruses. For complete details on the use and execution of this protocol, please refer to Hare et al. (2020). [Display omitted] •Single-cell assays are needed to measure IFN production in virus-infected cells•Immortalized THF cells with IFN-b reporter are used to measure IFN production•Individual cells are analyzed via flow cytometry and fluorescence microscopy•Simple single-cell assays can uncover data obscured in bulk population measurements Recent data suggest that cells respond to infection by upregulating the antiviral cytokine interferon-beta in a fraction of infected cells. Approaches are thus needed to study these responses on a single-cell level rather than bulk population. Here, we describe a protocol to analyze the IFN response of individual cells using flow cytometry and immunofluorescence microscopy. We show the heterogeneous IFN response to inactivated Sendai virus and human cytomegalovirus, but this protocol can be adapted to other viruses.