Rapid in vitro generation of bona fide exhausted CD8+ T cells is accompanied by Tcf7 promotor methylation
| Autor: | Leticia G. Leon, Yvonne M. Mueller, Inge Brouwers-Haspels, Marjan van Meurs, Eric M.J. Bindels, Ruben Boers, Joachim Boers, Peter D. Katsikis, Manzhi Zhao, Remco Hoogenboezem, Christopher J. Stairiker, Wilfred F. J. van IJcken, Stefan J. Erkeland, Jennifer L. Hope, Joost Gribnau, Caoimhe H. Kiernan |
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| Přispěvatelé: | Immunology, Developmental Biology, Cell biology, Hematology |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Physiology
CD8-Positive T-Lymphocytes Biochemistry Immune Receptors Mice White Blood Cells Animal Cells Immune Physiology Medicine and Health Sciences Lymphocytic choriomeningitis virus Cytotoxic T cell Hepatocyte Nuclear Factor 1-alpha Biology (General) Promoter Regions Genetic Innate Immune System 0303 health sciences DNA methylation Immune System Proteins T Cells Chemistry 030302 biochemistry & molecular biology hemic and immune systems Chromatin Cell biology Nucleic acids Cytokines Epigenetics Cellular Types DNA modification Chromatin modification Signal Transduction Research Article Chromosome biology QH301-705.5 Immune Cells Transgene Immunology Mice Transgenic Cytotoxic T cells chemical and pharmacologic phenomena Lymphocytic Choriomeningitis Microbiology complex mixtures 03 medical and health sciences In vivo Virology DNA-binding proteins Genetics Animals Gene Regulation Molecular Biology 030304 developmental biology Blood Cells T-cell receptor Biology and Life Sciences Proteins Cell Biology DNA Molecular Development RC581-607 In vitro Regulatory Proteins T Cell Receptors CTL Immune System Anergy Parasitology Gene expression Immunologic diseases. Allergy human activities CD8 Developmental Biology Transcription Factors |
| Zdroj: | PLoS Pathogens, Vol 16, Iss 6, p e1008555 (2020) PLoS Pathogens PLoS Pathogens (print), 16(6):e1008555. Public Library of Science |
| ISSN: | 1553-7374 1553-7366 |
| Popis: | Exhaustion is a dysfunctional state of cytotoxic CD8+ T cells (CTL) observed in chronic infection and cancer. Current in vivo models of CTL exhaustion using chronic viral infections or cancer yield very few exhausted CTL, limiting the analysis that can be done on these cells. Establishing an in vitro system that rapidly induces CTL exhaustion would therefore greatly facilitate the study of this phenotype, identify the truly exhaustion-associated changes and allow the testing of novel approaches to reverse or prevent exhaustion. Here we show that repeat stimulation of purified TCR transgenic OT-I CTL with their specific peptide induces all the functional (reduced cytokine production and polyfunctionality, decreased in vivo expansion capacity) and phenotypic (increased inhibitory receptors expression and transcription factor changes) characteristics of exhaustion. Importantly, in vitro exhausted cells shared the transcriptomic characteristics of the gold standard of exhaustion, CTL from LCMV cl13 infections. Gene expression of both in vitro and in vivo exhausted CTL was distinct from T cell anergy. Using this system, we show that Tcf7 promoter DNA methylation contributes to TCF1 downregulation in exhausted CTL. Thus this novel in vitro system can be used to identify genes and signaling pathways involved in exhaustion and will facilitate the screening of reagents that prevent/reverse CTL exhaustion. Author summary In this manuscript, we describe an in vitro method that rapidly establishes large numbers of exhausted CD8+ T cells. The exhaustion of CTL induced by this method has been fully validated by multiple approaches (cytokine production, polyfunctionality, cytotoxicity, in vivo proliferation, inhibitory receptors, transcription factors, RNAseq and DNA methylation). This method will facilitate not only the study of T cell exhaustion but also the screening of drugs. As proof of point, we use this method to show that TCF-1 downregulation in terminally exhausted T cells is accompanied by Tcf7 DNA promoter methylation and show that a transmethylase inhibitor can prevent TCF-1 downregulation. Our method presents a critical resource for the study of CTL exhaustion and the screening of drugs and interventions. |
| Databáze: | OpenAIRE |
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