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Human defensin 5 is stored in precursor form in normal Paneth cells and is expressed by some villous epithelial cells and by metaplastic Paneth cells in the colon in inflammatory bowel disease
  1. R N Cunliffea,
  2. F R A J Rosea,
  3. J Keyteb,
  4. L Abberleya,
  5. W C Chanc,
  6. Y R Mahidaa
  1. aDivision of Gastroenterology, University of Nottingham, UK, bDepartment of Biochemistry, University of Nottingham, UK, cDepartment of Pharmaceutical Sciences, University of Nottingham, UK
  1. Y R Mahida, Division of Gastroenterology, University Hospital, Queen's Medical Centre, Nottingham NG7 2UH, UK. Yash.Mahida{at}nottingham.ac.uk

Abstract

BACKGROUND AND AIMS Intestinal epithelial cell derived antimicrobial peptides of the defensin family may play a major role in host defence against microorganisms. Our aims were to (i) isolate, characterise, and investigate the processing of human defensin 5 (HD-5) in normal Paneth cells and (ii) investigate expression of HD-5 in active inflammatory bowel disease (IBD).

METHODS Antiserum raised against chemically synthesised putative mature HD-5 was used for immunohistochemistry and purification of HD-5 from extracts of normal terminal ileal crypts.

RESULTS In normal and Crohn's disease terminal ileum, HD-5 immunoreactivity was seen in Paneth cells and in some villous epithelial cells. Normal colonic mucosa did not express HD-5 but HD-5 immunoreactivity was seen in cells in the colonic crypt region of many IBD samples.N-terminal amino acid sequence analysis of HD-5 purified from normal terminal ileal Paneth cells consistently showed the predicted sequence of the precursor form of the peptide. Following stimulation of isolated intact normal terminal ileal crypts, a truncated form of HD-5, with theN-terminal sequence GEDNQLAIS, was detected in the supernatant.

CONCLUSIONS (i) HD-5 is present only in the precursor form in normal terminal ileal Paneth cells and is processed to the mature form during and/or after secretion, (ii) some villous epithelial cells express HD-5, and (iii) HD-5 is expressed by metaplastic Paneth cells in the colon in IBD.

  • inflammatory bowel disease
  • human defensin 5
  • Paneth cells
  • Abbreviations used in this paper

    IBD
    inflammatory bowel disease
    HD-5
    HD-6, human defensins 5 and 6
    RP-HPLC
    reverse phase-high performance liquid chromatography
    TFA
    trifluoroacetic acid
    PBS
    phosphate buffered saline
    ITF
    intestinal trefoil factor
    HBSS
    Hanks balanced salt solution
    LPS
    lipopolysaccharide
    SDS-PAGE
    sodium dodecyl sulphate-polyacrylamide gel electrophoresis
    PVDF
    polyvinylidene difluoride
    TTBS
    Tris-Tween buffered saline
    CFU
    colony forming unit
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  • Abbreviations used in this paper

    IBD
    inflammatory bowel disease
    HD-5
    HD-6, human defensins 5 and 6
    RP-HPLC
    reverse phase-high performance liquid chromatography
    TFA
    trifluoroacetic acid
    PBS
    phosphate buffered saline
    ITF
    intestinal trefoil factor
    HBSS
    Hanks balanced salt solution
    LPS
    lipopolysaccharide
    SDS-PAGE
    sodium dodecyl sulphate-polyacrylamide gel electrophoresis
    PVDF
    polyvinylidene difluoride
    TTBS
    Tris-Tween buffered saline
    CFU
    colony forming unit
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