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INFLAMMATION AND INFLAMMATORY BOWEL DISEASE |
(MIP-3) production in normal colon and inflammatory bowel disease
1 Division of Gastroenterology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston MA 02215, USA
2 Immunobiology Center, Mount Sinai School of Medicine, New York, NY 10029, USA
Correspondence to:
Correspondence to:
Dr A C Keates, Dana 501, Gastroenterology, Beth Israel Deaconess Medical Center, 330 Brookline Avenue, Boston, MA 02215, USA;
akeates{at}caregroup.harvard.edu
ABSTRACT
Background and aim: Macrophage inflammatory protein 3
(MIP-3) is a recently described lymphocyte directed C-C chemokine expressed predominately at extralymphoid sites, including the intestine. The aim of this study was to determine whether colonic epithelial cells produce MIP-3 and whether its expression is upregulated in inflammatory bowel disease.
Methods and results: We found that interleukin 1ß and tumour necrosis factor dose dependently stimulated MIP-3 production in Caco-2 and HT-29 intestinal epithelial cells. In cytokine treated Caco-2 and HT-29 cells, a significant increase in MIP-3 protein production was observed after three hours and continued for at least 24 hours. Analysis of colonic tissues by quantitative real time polymerase chain reaction and ELISA revealed significantly elevated MIP-3 mRNA levels (7.9-fold; p<0.05) and protein levels (8.9-fold; p<0.05) in Crohn’s disease compared with controls or ulcerative colitis. MIP-3 immunoreactivity in normal colon and inflammatory bowel disease was principally associated with crypt and surface epithelial cells. Moreover, MIP-3 protein levels were elevated in primary epithelial cells isolated from patients with inflammatory bowel disease.
Conclusions: These findings indicate that increased enterocyte MIP-3 production may play an important role in lymphocyte activation and recruitment to the colonic epithelium in Crohn’s disease and ulcerative colitis.
Keywords: chemokine; inflammation; enterocyte; intestine
Abbreviations: MIP, macrophage inflammatory protein; MCP-1, monocyte chemotactic protein 1; MAdCAM-1, mucosal addressin cell adhesion molecule 1; IL, interleukin; TNF-, tumour necrosis factor ; IBD, inflammatory bowel disease; PBS, phosphate buffered saline; BSA, bovine serum albumin; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; RT-PCR, reverse transcription-polymerase chain reaction; TBS, Tris buffered saline
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