title: Cell-Based Screen Identifies Human Interferon-Stimulated Regulators of Listeria monocytogenes Infection.
abstract: The type I interferon (IFN) activated transcriptional response is a critical antiviral defense mechanism, yet its role in bacterial pathogenesis remains less well characterized. Using an intracellular pathogen Listeria monocytogenes (Lm) as a model bacterial pathogen, we sought to identify the roles of individual interferon-stimulated genes (ISGs) in context of bacterial infection. Previously, IFN has been implicated in both restricting and promoting Lm growth and immune stimulatory functions in vivo. Here we adapted a gain-of-function flow cytometry based approach to screen a library of more than 350 human ISGs for inhibitors and enhancers of Lm infection. We identify 6 genes, including UNC93B1, MYD88, AQP9, and TRIM14 that potently inhibit Lm infection. These inhibitors act through both transcription-mediated (MYD88) and non-transcriptional mechanisms (TRIM14). Further, we identify and characterize the human high affinity immunoglobulin receptor FcγRIa as an enhancer of Lm internalization. Our results reveal that FcγRIa promotes Lm uptake in the absence of known host Lm internalization receptors (E-cadherin and c-Met) as well as bacterial surface internalins (InlA and InlB). Additionally, FcγRIa-mediated uptake occurs independently of Lm opsonization or canonical FcγRIa signaling. Finally, we established the contribution of FcγRIa to Lm infection in phagocytic cells, thus potentially linking the IFN response to a novel bacterial uptake pathway. Together, these studies provide an experimental and conceptual basis for deciphering the role of IFN in bacterial defense and virulence at single-gene resolution.
authors.affiliation: Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, Texas, United States of America.
authors.forename: Sofya S
authors.identifier: http://orcid.org/0000-0001-9706-8156
authors.initials: SS
authors.lastname: Perelman
authors.affiliation: Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, Texas, United States of America.
authors.forename: Michael E
authors.identifier: http://orcid.org/0000-0001-5493-0965
authors.initials: ME
authors.lastname: Abrams
authors.affiliation: Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, Texas, United States of America.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Eitson
authors.affiliation: Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, Texas, United States of America.
authors.forename: Didi
authors.initials: D
authors.lastname: Chen
authors.affiliation: Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, Texas, United States of America.
authors.forename: Alyssa
authors.initials: A
authors.lastname: Jimenez
authors.affiliation: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas, United States of America.
authors.forename: Marcel
authors.initials: M
authors.lastname: Mettlen
authors.affiliation: Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, Texas, United States of America.
authors.forename: John W
authors.initials: JW
authors.lastname: Schoggins
authors.affiliation: Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, Texas, United States of America.
authors.forename: Neal M
authors.initials: NM
authors.lastname: Alto
mesh_terms: D002460:Cell Line; D005434:Flow Cytometry; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D015151:Immunoblotting; D007370:Interferon Type I / Q000276:immunology*; D008089:Listeria monocytogenes / Q000276:immunology; D008088:Listeriosis / Q000235:genetics / Q000276:immunology*; D008855:Microscopy, Electron, Scanning; D016133:Polymerase Chain Reaction; D059467:Transcriptome; D014774:Virulence / Q000276:immunology*
chemical_list: D007370:Interferon Type I