Article Text
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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