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. 1998 Feb;42(2):214–219. doi: 10.1136/gut.42.2.214

Expression of interleukin 1β and interleukin 1β converting enzyme by intestinal macrophages in health and inflammatory bowel disease

M McAlindon 1, C Hawkey 1, Y Mahida 1
PMCID: PMC1726995  PMID: 9536946

Abstract

Background—In the lipopolysaccharide (LPS) stimulated peripheral blood monocyte, the precursor form of interleukin 1β (IL-1β, 31 kD) is processed by IL-1β converting enzyme (ICE) to the mature, bioactive form (17 kD). IL-1β is a proinflammatory cytokine which is likely to have a role in the pathogenesis of inflammatory bowel disease (IBD). 
Aims—To investigate the expression and processing of IL-1β and ICE by tissue macrophages from normal and IBD colonic mucosa. 
Methods—Mucosal biopsy specimens and lamina propria cells from normal and IBD colons were studied by reverse transcription polymerase chain reaction (RT-PCR), western blot analysis, and ELISA (enzyme linked immunosorbent assay). 
Results—Normal colonic macrophages synthesised only the precursor form of IL-1β whereas in IBD the mature form was also produced. Similarly, cells from normal colonic mucosa synthesised ICE as the precursor (p45) only, whereas macrophages from IBD colons produced active (p20) ICE. Ac-Tyr-Val-Ala-Asp-CHO, a specific peptide aldehyde inhibitor of ICE, significantly reduced the amount of mature IL-1β released by isolated IBD macrophages (from a median of 1.2 (range 0.78-4.42) ng/ml to 0.43 (0.21-1.6) ng/ml; p<0.01). 
Conclusions—Exposure of normal colonic macrophages to LPS only induces the production of the precursor form of IL-1β, because the cells fail to activate ICE. In contrast, IBD colonic macrophages are able to activate ICE and hence release mature IL-1β in a manner similar to circulating monocytes. This is consistent with IBD macrophages being recently recruited from the circulating monocyte population. Targeted inhibition of ICE may represent a novel form of therapy in IBD. 



Keywords: interleukin 1β; interleukin 1β converting enzyme; macrophages; lipopolysaccharide; ulcerative colitis; Crohn's disease

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Figure 1 .

Figure 1

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Expression of mRNA transcripts for IL-1β and glyceraldehyde-6-phosphate dehydrogenase (GAP) in a normal colonic biopsy specimen; lamina propria mononuclear cells (MNC) isolated from samples of normal colonic mucosa; and a biopsy specimen with active IBD. The figure shows an example of 10/13 normal colonic biopsy samples which did not express transcripts for IL-1β. 


Figure 2 .

Figure 2

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Expression of IL-1β protein in LPS stimulated peripheral blood mononuclear cell culture supernatants in the absence (-) and presence (+) of the ICE inhibitor, Ac-Tyr-Val-Ala-Asp-CHO. 


Figure 3 .

Figure 3

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Expression of IL-1β protein by LPS stimulated lamina propria cells isolated from normal and IBD colonic mucosa. Cell lysates and supernatants were studied by western blot analysis. In cells obtained from IBD colon, both 31 kD and 17 kD forms of the cytokine were detected in the supernatants. The figure shows representative experiments performed on lamina propria cells obtained from four normal and five IBD colons. 


Figure 4 .

Figure 4

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Expression of ICE and GAPDH mRNA in biopsy specimens from normal and IBD mucosa. 


Figure 5 .

Figure 5

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Expression of ICE protein by LPS stimulated lamina propria cells isolated from normal and IBD colonic mucosa. In contrast to IBD, the p20 form of ICE was not present in the lysate of cells from normal colonic mucosa. The lighter band between p45 and p20 is likely to represent a 36 kD intermediate cleavage product of p45 (and which is further processed down to p20).38


Figure 6 .

Figure 6

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Release of IL-1β protein by IBD lamina propria cells cultured in the presence and absence of ICE inhibitor. 


Figure 7 .

Figure 7

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ICE inhibitor (Ac-Tyr-Val-Ala-Asp-CHO) reduces release of mature IL-1β by lamina propria cells isolated from mucosa with active IBD. Paired samples (n=7) of isolated lamina propria cells were cultured with LPS, in the presence or absence of Ac-Tyr-Val-Ala-Asp-CHO.


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