1. An Image-Based Genetic Assay Identifies Genes in T1D Susceptibility Loci Controlling Cellular Antiviral Immunity in Mouse
- Author
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Hermann Bauer, Kara G. Lassen, Gautam Goel, Harry Sokol, Ramnik J. Xavier, Juan Liao, Aivi Doan, Humberto Jijon, Bernhard G. Herrmann, Ira R. Kim, Université Pierre et Marie Curie - Paris 6 (UPMC), Service de Gastroentérologie et nutrition [CHU Saint-Antoine], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Saint-Antoine [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU), CHU Saint-Antoine [AP-HP], and Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)
- Subjects
Interleukin-27 ,lcsh:Medicine ,Autoimmunity ,Herpesvirus 1, Human ,Virus Replication ,Biochemistry ,Immune Receptors ,Intracellular Receptors ,Mice ,Signal Initiation ,RNA interference ,Cell Signaling ,Molecular Cell Biology ,Gene Regulatory Networks ,RNA, Small Interfering ,lcsh:Science ,Immune Response ,Genetic Interference ,Genetics ,Immunity, Cellular ,Multidisciplinary ,Immune System Proteins ,[CHIM.ORGA]Chemical Sciences/Organic chemistry ,GTPase-Activating Proteins ,Mechanisms of Signal Transduction ,Infectious Disease Immunology ,Recombinant Proteins ,3. Good health ,Cytokines ,Epigenetics ,Disease Susceptibility ,Chemokines ,Research Article ,Signal Transduction ,Feedback Regulation ,Immunology ,Biology ,Genetic Predisposition ,Proinflammatory cytokine ,Cell Line ,Interferon-gamma ,Immune system ,Genetic predisposition ,Animals ,Gene Disruption ,Molecular Biology Techniques ,Molecular Biology ,Inflammation ,Molecular Biology Assays and Analysis Techniques ,Biology and life sciences ,Macrophages ,lcsh:R ,Proteins ,Computational Biology ,Interferon-beta ,Cell Biology ,Virology ,High Throughput Screening ,High-Throughput Screening Assays ,RNA interference screens ,Diabetes Mellitus, Type 1 ,Viral replication ,Microscopy, Fluorescence ,Genetic Loci ,Genetics of Disease ,lcsh:Q ,Interferon Regulatory Factor-3 ,Clinical Immunology ,Gene Function ,IRF3 ,Interferon regulatory factors ,Genetic screen - Abstract
International audience; The pathogenesis of complex diseases, such as type 1 diabetes (T1D), derives from interactions between host genetics and environmental factors. Previous studies have suggested that viral infection plays a significant role in initiation of T1D in genetically predisposed individuals. T1D susceptibility loci may therefore be enriched in previously uncharacterized genes functioning in antiviral defense pathways. To identify genes involved in antiviral immunity, we performed an image-based high-throughput genetic screen using short hairpin RNAs (shRNAs) against 161 genes within T1D susceptibility loci. RAW 264.7 cells transduced with shRNAs were infected with GFP-expressing herpes simplex virus type 1 (HSV-1) and fluorescent microscopy was performed to assess the viral infectivity by fluorescence reporter activity. Of the 14 candidates identified with high confidence, two candidates were selected for further investigation, Il27 and Tagap. Administration of recombinant IL-27 during viral infection was found to act synergistically with interferon gamma (IFN-γ) to activate expression of type I IFNs and proinflammatory cytokines, and to enhance the activities of interferon regulatory factor 3 (IRF3). Consistent with a role in antiviral immunity, Tagap-deficient macrophages demonstrated increased viral replication, reduced expression of proinflammatory chemokines and cytokines, and decreased production of IFN-β. Taken together, our unbiased loss-of-function genetic screen identifies genes that play a role in host antiviral immunity and delineates roles for IL-27 and Tagap in the production of antiviral cytokines.
- Published
- 2014