RNA interference screen in primary human T cells reveals FLT3 as a modulator of IL-10 levels

Anne L Astier, Gaëlle Beriou, Thomas M Eisenhaure, Stephen M Anderton, David A Hafler, Nir Hacohen

Research output: Contribution to journalArticlepeer-review

Abstract

Functional studies of human primary immune cells have been hampered by the lack of tools to silence gene functions. In this study, we report the application of a lentiviral RNA interference library in primary human T cells. Using a subgenomic short hair RNA library targeting approximately 1000 signaling genes, we identified novel genes that control the levels of IL-10 produced. IL-10 is a potent anti-inflammatory cytokine secreted by several cell types, including T regulatory type 1 cells, a subset of T regulatory cells that exert their suppressive activity through IL-10 secretion. FLT3, a known hematopoeitic growth factor, was found to be a negative regulator of IL-10 levels in activated T cells. This was based on several observations. First, FLT3 and its ligand (FL) were both induced by T cell activation. Second, silencing of FLT3 led to increased IL-10 levels, whereas addition of FL suppressed IL-10 secretion and increased FLT3 surface levels. Third, engagement of CD46, a known inducer of T regulatory type 1 cells, upregulated surface FLT3, and secreted FL, which then inhibited IL-10 production by T cells. Hence, FL and FLT3 form a novel regulatory feedback loop that limits IL-10 production in T cells. Our results identified FLT3 as a new regulator of T cell function and offer a strategy to genetically dissect specific pathways in T cells.
Original languageEnglish
Pages (from-to)685-93
Number of pages9
JournalJournal of Immunology
Volume184
Issue number2
DOIs
Publication statusPublished - 2010

Keywords

  • Antigens, CD46
  • Cells, Cultured
  • Feedback, Physiological
  • Gene Expression Regulation
  • Gene Library
  • Humans
  • Immunologic Factors
  • Interleukin-10
  • Lentivirus
  • Ligands
  • Lymphocyte Activation
  • RNA Interference
  • RNA, Small Interfering
  • T-Lymphocytes
  • fms-Like Tyrosine Kinase 3

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