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Clinical & Experimental Allergy

Transforming growth factor-bβ1 mediates coexpression of the integrin subunit aαE and the chymase mouse mast cell protease-1 during the early differentiation of bone marrow-derived mucosal mast cell homologues

Authors

  • S. H. Wright,

    1. Department of Veterinary Clinical Studies and The Wellcome Trust Centre For Research in Comparative Respiratory Medicine, Royal (Dick) School of Veterinary Studies, The University of Edinburgh, Easter Bush Veterinary Centre, Easter Bush, Roslin
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  • J. Brown,

    1. Department of Veterinary Clinical Studies and The Wellcome Trust Centre For Research in Comparative Respiratory Medicine, Royal (Dick) School of Veterinary Studies, The University of Edinburgh, Easter Bush Veterinary Centre, Easter Bush, Roslin
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  • P. A. Knight,

    1. Department of Veterinary Clinical Studies and The Wellcome Trust Centre For Research in Comparative Respiratory Medicine, Royal (Dick) School of Veterinary Studies, The University of Edinburgh, Easter Bush Veterinary Centre, Easter Bush, Roslin
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  • E. M. Thornton,

    1. Department of Veterinary Clinical Studies and The Wellcome Trust Centre For Research in Comparative Respiratory Medicine, Royal (Dick) School of Veterinary Studies, The University of Edinburgh, Easter Bush Veterinary Centre, Easter Bush, Roslin
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  • P. J. Kilshaw,

    1. Department of Immunology, The Babraham Institute, Babraham Hall, Babraham, Cambridge, UK
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  • H. R. P. Miller

    Corresponding author
    1. Department of Veterinary Clinical Studies and The Wellcome Trust Centre For Research in Comparative Respiratory Medicine, Royal (Dick) School of Veterinary Studies, The University of Edinburgh, Easter Bush Veterinary Centre, Easter Bush, Roslin
      Hugh R. P. Miller, Department of Veterinary Clinical Studies and The Wellcome Trust Centre For Research in Comparative Respiratory Medicine, Royal (Dick) School of Veterinary Studies, The University of Edinburgh, Easter Bush Veterinary Centre, Easter Bush, Roslin, Midlothian, EH25 9RG, UK. E-mail: Hugh.Miller@ed.ac.uk
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Hugh R. P. Miller, Department of Veterinary Clinical Studies and The Wellcome Trust Centre For Research in Comparative Respiratory Medicine, Royal (Dick) School of Veterinary Studies, The University of Edinburgh, Easter Bush Veterinary Centre, Easter Bush, Roslin, Midlothian, EH25 9RG, UK. E-mail: Hugh.Miller@ed.ac.uk

Abstract

Background

Mucosal mast cells (MMC) play a central role in gut hypersensitivities and inflammation. They are morphologically, biochemically and functionally distinct from their connective tissue counterparts. Massive hyperplasia of MMC occurs 7–10 days after intestinal infection with nematodes but it has never been possible to replicate this phenomenon in vitro.

Objective

(1) To determine whether mouse bone marrow-derived mast cells (mBMMC) grown in the presence of transforming growth factor (TGF)-β1 could develop over the same time frame (7–10 days) as MMC in parasitized mice. (2) To compare the early expression of surface receptors (integrins αE and β7, c-kit and FcεR) with that of the MMC-specific granule chymase mouse mast cell protease-1 (mMCP-1).

Methods

Mouse bone marrow cells were cultured in the presence of IL-9, IL-3 and Stem Cell Factor (SCF) with or without TGF-β1. mBMMC were quantified after toluidine blue or Leishmans' staining. Expression of MMC-specific mouse mast cell proteases was analysed by ELISA, immunohistochemistry and RT-PCR. Surface antigen expression was characterized by flow cytometry and confocal microscopy.

Results

TGF-β1 promotes the development of abundant MMC-like mBMMC from bone marrow progenitor cells with kinetics, which closely parallel that seen in vivo. mRNA transcripts encoding mMCP-1 and -2 are readily detectable by day 4 ex vivo in cultures grown in the presence of TGF-β1. Between 30 and 40% and 75–90% of the cells in these cultures on days 4 and 7, respectively, have typical mast cell morphology, are c-kit+, FcεR+, integrin αEβ7+, and express and secrete abundant mMCP-1. The integrin αE subunit is coexpressed with mMCP-1.

Conclusion

The kinetics of mMCP-1+E+ mBMMC development, regulated by TGF-β1, are consistent with that seen in vivo in the parasitized intestine. The normally down-regulatory functions of TGF-β1 in haematopoiesis are superseded in this culture system by its ability to promote the early expression of αE and mMCP-1.

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