RT Journal Article
SR Electronic
T1 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
JF Gut
JO Gut
FD BMJ Publishing Group Ltd and British Society of Gastroenterology
SP 176
OP 185
DO 10.1136/gut.48.2.176
VO 48
IS 2
A1 R N Cunliffe
A1 F R A J Rose
A1 J Keyte
A1 L Abberley
A1 W C Chan
A1 Y R Mahida
YR 2001
UL http://gut.bmj.com/content/48/2/176.abstract
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