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Autoregulation of Th1-mediated inflammation by twist1
Uwe Niesner, Inka Albrecht, Marko Janke, Cornelia Doebis, Christoph Loddenkemper, Maria H. Lexberg, Katharina Eulenburg, Stephan Kreher, Juliana Koeck, Ria
Baumgrass, Kerstin Bonhagen, Thomas Kamradt, Philipp Enghard, Jens Y. Humrich, Sascha Rutz, Ulf Schulze-Topphoff, Orhan Aktas, Sina Bartfeld, Helena Radbruch, Ahmed N. Hegazy, Max Löhning, Daniel C. Baumgart, Rainer Duchmann, Martin
Rudwaleit, Thomas Häupl, Inna Gitelman, Veit Krenn, Joachim Gruen, Jochen Sieper, Martin Zeitz, Bertram Wiedenmann, Frauke Zipp, Alf Hamann, Michal Janitz,
Alexander Scheffold, Gerd R. Burmester, Hyun D. Chang, and Andreas Radbruch
Journal of Experimental MedicineVolume 205(8):1889-1901
August 2, 2008
© 2008 Niesner et.al.
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Th cell balance
naive T cell
TGFβIL-1/IL-6/IL-23
IFNγ
IL-17A-FIL-22
IL-4IL-10IL-13IL-5
Activation STAT4Production IFNγActivation Tbet
Activation RORC2
Activation STAT6Activation GATA3
IL-4
IL-12
Th1
Th17
Th2
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Twist1: a basic helix-loop-helix transcription factor
basic helix-loop-helix transcription factorbinds to a consensus sequence called e-box: CANNTG
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Transcriptional regulation of bHLH transcription factors
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Repeated Th1 stimulation
naive CD4+ CD62L+ T cellsfrom DO11.10 mice (carrying a TCR specific for Ovalbumin)
splenic APCOvalbuminIL-12anti-IL-4
6d
restimulation
gene array
...
38 fold upregulation of twist1 expression in 4 times restimulated Th1 cells
PMA/iono CD3/CD28
quantitative real-time PCR
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Twist1 expression is induced in Th1, but not Th2 or Th17 cells
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Twist1 induction requires IL-12 signaling via STAT4, but not IFN-γ or T-bet
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Signaling through NFAT and NF-κB is required for induction of twist1 expression
PDTC blocksNF-κBBTP blocks NFATCSA blocks IL-2 signaling
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Ex vivo isolated memory Th cells express twist1
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Generation of Th1 cells constitutively expressing twist1
• d0: splenic DO11.10 cells activated in vitro
• d2: infection with twist1 encoding or control retrovirus
• d5: sort of GFP+
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Genes differentially expressed upon ectopic twist1 overexpression
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Twist1 suppresses the expression of effector cytokines
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NF-κB driven luciferase reporter
• d0: splenic DO11.10 cells activated in vitro
• d2: infection with twist1 or I-κBαM encoding retrovirus
• d3: nucleoporation with a mixture of luciferase encoding plasmids
• d5: sort of GFP+
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Inhibition of NF-κB–mediated signaling by twist1 is promoter-specific
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DTH model
• 6d-old-Th1 cells overexpressing twist1 injected i.v. in BALB/c
• 1d later OVA + IFA injection in footpad
• messure of swelling of the footpad
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Ectopic twist1 overexpression controls DTH
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Twist1 knockdown increases inflammatory response in murine arthritis
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Conclusions
• repeatedly stimulated Th1 cells express twist1• twist1 expression is NF-κB, NFAT and STAT4
dependent• twist1 is expressed in humans and mice
– in Th1 effector memory cells – on CD4+ T cells of inflammed tissue
• Tbet is not needed for twist1 expression• twist1 binds to e-boxes in the regulatory regions
of specific target genes• twist1 reduces activation-induced expression IL-2,
IFN-γ, TNF-α on the protein level and theirby dampening an inflammation