PT  - JOURNAL ARTICLE
AU  - Cunliffe, R N
AU  - Rose, F R A J
AU  - Keyte, J
AU  - Abberley, L
AU  - Chan, W C
AU  - Mahida, Y R
TI  - 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
AID  - 10.1136/gut.48.2.176
DP  - 2001 Feb 01
TA  - Gut
PG  - 176--185
VI  - 48
IP  - 2
4099  - http://gut.bmj.com/content/48/2/176.short
4100  - http://gut.bmj.com/content/48/2/176.full
SO  - Gut2001 Feb 01; 48
AB  - 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.IBDinflammatory bowel diseaseHD-5HD-6, human defensins 5 and 6RP-HPLCreverse phase-high performance liquid chromatographyTFAtrifluoroacetic acidPBSphosphate buffered salineITFintestinal trefoil factorHBSSHanks balanced salt solutionLPSlipopolysaccharideSDS-PAGEsodium dodecyl sulphate-polyacrylamide gel electrophoresisPVDFpolyvinylidene difluorideTTBSTris-Tween buffered salineCFUcolony forming unit