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Blockade of transforming growth factor β upregulates T-box transcription factor T-bet, and increases T helper cell type 1 cytokine and matrix metalloproteinase-3 production in the human gut mucosa
  1. A Di Sabatino1,
  2. K M Pickard2,
  3. D Rampton3,
  4. L Kruidenier4,
  5. L Rovedatti5,
  6. N A B Leakey1,
  7. G R Corazza5,
  8. G Monteleone6,
  9. T T MacDonald1
  1. 1 Centre for Infectious Disease, Institute of Cell and Molecular Science, Barts and the London School of Medicine and Dentistry, London, UK
  2. 2 Division of Infection, Inflammation and Repair, University of Southampton School of Medicine, Southampton, UK
  3. 3 Centre for Gastroenterology, Institute of Cell and Molecular Science, Barts and the London School of Medicine and Dentistry, London, UK
  4. 4 GI and Neurosciences CEDD, GlaxoSmithKline Ltd, Harlow, UK
  5. 5 First Department of Medicine, Fondazione IRCCS Policlinico S. Matteo, Centro per lo Studio e la Cura delle Malattie Infiammatorie Croniche Intestinali, University of Pavia, Pavia, Italy
  6. 6 Dipartimento di Medicina Interna e Centro di Eccellenza per lo Studio delle Malattie Complesse e Multifattoriali, Universitè Tor Vergata, Rome, Italy
  1. Professor T T MacDonald, Centre for Infectious Disease, Institute of Cell and Molecular Science, Barts and the London School of Medicine and Dentistry, Whitechapel, London E1 2AT, UK; t.t.macdonald{at}qmul.ac.uk

Abstract

Background and Aims: The role of transforming growth factor β (TGFβ) in inhibiting T cell function in the normal gut has been studied in animal models. However, the impact of TGFβ inhibition on T cells in the normal human gut remains poorly understood. The effect of TGFβ blockade in normal intestinal biopsies grown ex vivo and lamina propria mononuclear cells (LPMCs) on T-bet, a T-box transcription factor required for T helper cell type (Th)1 differentiation, interferon γ (IFNγ) production, T cell apoptosis and matrix metalloproteinase (MMP)-3 production has therefore been tested.

Methods: TGFβ transcripts were determined by quantitative reverse transcription-PCR in laser-captured gut epithelium and lamina propria. Biopsies and LPMCs were cultured with anti-TGFβ neutralising antibody. After 24 h culture, T-bet was determined by immunoblotting, and T cell apoptosis was assessed by flow cytometry. IFNγ, tumour necrosis factor α (TNFα), interleukin (IL) 2, IL6, IL8, IL10, IL12p70 and IL17 were measured by ELISA. MMP-3 and tissue inhibitor of matrix metalloproteinase (TIMP)-1 were assessed by immunoblotting.

Results: A higher number of TGFβ transcripts was found in the lamina propria than in the epithelium in normal gut. T-bet expression was significantly higher in biopsies and LPMCs cultured with anti-TGFβ antibody than in those cultured with control antibody. TGFβ blockade downregulated T cell apoptosis, and induced a significant increase in IFNγ, TNFα, IL2, IL6, IL8 and IL17 production. A higher expression of MMP-3, but not TIMP-1, was observed in the tissue and supernatant of biopsies treated with anti-TGFβ antibody.

Conclusions: The findings support a crucial role for TGFβ in dampening T cell-mediated tissue-damaging responses in the human gut.

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Footnotes

  • Competing interests: None.

  • Patient consent: Consent was obtained from all subjects.

  • Ethics approval: Ethics approval was obtained from the local Ethics Committee.

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