Colonic epithelial cells are a major site of macrophage inflammatory protein 3alpha (MIP-3alpha) production in normal colon and inflammatory bowel disease

Abstract

Background and aim: Macrophage inflammatory protein 3α (MIP-3α) 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-3α and whether its expression is upregulated in inflammatory bowel disease. Methods and results: We found that interleukin 1β and tumour necrosis factor α dose dependently stimulated MIP-3α production in Caco-2 and HT-29 intestinal epithelial cells. In cytokine treated Caco-2 and HT-29 cells, a significant increase in MIP-3α 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-3α 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-3α immunoreactivity in normal colon and inflammatory bowel disease was principally associated with crypt and surface epithelial cells. Moreover, MIP-3α protein levels were elevated in primary epithelial cells isolated from patients with inflammatory bowel disease. Conclusions: These findings indicate that increased enterocyte MIP-3α production may play an important role in lymphocyte activation and recruitment to the colonic epithelium in Crohn's disease and ulcerative colitis.

ESE-1, an Enterocyte-specific Ets Transcription Factor, Regulates MIP-3α Gene Expression in Caco-2 Human Colonic Epithelial Cells

“…This stimulus was selected because colonic levels of this pro-inflammatory cytokine are significantly elevated in human IBD (28). Furthermore, we have shown previously that IL-1␤ can dose-dependently upregulate MIP-3␣ mRNA and protein production in Caco-2 colonic epithelial cells (15).…”

“…Promoter Region-Recent studies from our laboratory have shown that enterocytes are a major site of MIP-3␣ production in human colon and that epithelial MIP-3␣ protein levels are elevated in IBD (15). However, the molecular mechanisms that control MIP-3␣ gene expression in intestinal epithelial cells are poorly understood.…”

“…Moreover, in a recent study we have shown that MIP-3␣ production by cytokine-stimulated Caco-2 cells requires de novo mRNA synthesis (15). To begin to characterize the areas of the 5Ј promoter region that regulate MIP-3␣ gene expression, a series of luciferase reporter constructs containing sequentially truncated fragments of the MIP-3␣ promoter were created and transfected into Caco-2 intestinal epithelial cells (Fig.…”

“…MIP-3␣ also induces T-lymphocyte adhesion to the gastrointestinal-specific vascular addressin MAd-CAM-1 (13). In recent studies we, and others, have shown that enterocytes are a major site of MIP-3␣ production in human colon and that epithelial MIP-3␣ protein levels are elevated in IBD 1 (15,16). These studies are complemented by a recent report that MIP-3␣ mRNA levels were significantly elevated in chronically inflamed colons from IL-10 knockout mice (17).…”

Stromal CCR6 drives tumor growth in a murine transplantable colon cancer through recruitment of tumor-promoting macrophages

“…1 It is well known that CCL20 can be produced by intestinal epithelial cells, particularly in response to stress. 2,3 CCL20-CCR6 interactions have recently been shown to be instrumental in intestinal tumorigenesis and colorectal cancer growth. [4][5][6] While the direct role of CCL20-CCR6 interactions on promoting malignant behavior in neoplastic colon cancer cells has been well demonstrated, [6][7][8] despite the fact that chemokines are most known for their effects on immune cells, the role of these interactions on immune cells in colon cancer stroma has not been well studied.…”