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. 1999 Nov;45(5):705–712. doi: 10.1136/gut.45.5.705

Dexamethasone inhibition of leucocyte adhesion to rat mesenteric postcapillary venules: role of intercellular adhesion molecule 1 and KC

A Tailor 1, A Tomlinson 1, A Salas 1, J Panes 1, D Granger 1, R Flower 1, M Perretti 1
PMCID: PMC1727732  PMID: 10517906

Abstract

BACKGROUND—A previous study showed that the glucocorticoid dexamethasone, at doses of 100 µg/kg and above, inhibited leucocyte adhesion to rat mesenteric postcapillary venules activated with interleukin 1β (IL-1β), as assessed by videomicroscopy.
AIMS—To identify whether the adhesion molecule, intercellular adhesion molecule 1 (ICAM-1), or the chemokine KC could be targeted by the steroid to mediate its antiadhesive effect.
METHODS—Rat mesenteries were treated with IL-1β (20 ng intraperitoneally) and the extent of leucocyte adhesion measured at two and four hours using intravital microscopy. Rats were treated with dexamethasone, and passively immunised against ICAM-1 or KC. Endogenous expression of these two mediators was validated by immunohistochemistry, ELISA, and the injection of specific radiolabelled antibodies.
RESULTS—Dexamethasone greatly reduced IL-1β induced leucocyte adhesion, endothelial expression of ICAM-1 in the postcapillary venule, and release of the mast cell derived chemokine KC. Injection of specific antibodies to the latter mediators was also extremely effective in downregulating (>80%) IL-1β induced leucocyte adhesion.
CONCLUSIONS—Induction by IL-1β of endogenous ICAM-1 and KC contributes to leucocyte adhesion to inflamed mesenteric vessels. Without excluding other possible mediators, these data clearly show that dexamethasone interferes with ICAM-1 expression and KC release from mast cells, resulting in suppression of leucocyte accumulation in the bowel wall, which is a prominent feature of several gastrointestinal pathologies.


Keywords: inflammation; glucocorticoids; intravital microscopy; mast cell; neutrophil; endothelium

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Figure 1  .

Figure 1  

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Effect of dexamethasone (DEX) on interleukin (IL) 1β induced leucocyte adhesion and emigration in rat mesenteric postcapillary venules. Data are mean (SEM); n=6 rats. *p<0.05 v respective IL-1β group.

Figure 2  .

Figure 2  

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Involvement of intercellular adhesion molecule (ICAM) 1 in interleukin (IL) 1β induced cell adhesion and emigration. (A) Animals were treated with 2 mg/kg mouse IgG or mouse antirat ICAM-1 monoclonal antibody one hour prior to IL-1β administration (20 ng intraperitoneally). Mesenteries were exposed two hours later, and the degree of cell adhesion (per 100 µm vessel wall) and emigration quantified by videomicroscopy. Data are mean (SEM); n=6 rats. *p<0.05 v mouse IgG group. (B) ICAM-1 expression in the mesenteric tissue as assessed by the dual antibody technique. Rats were treated with rat IL-1β (20 ng intraperitoneally) and some of them pretreated with dexamethasone (DEX) one hour prior to the cytokine. At the reported times post-IL-1β, the specific radioactivity due to endogenous ICAM-1 was determined. Data are mean (SEM); n=5 rats per group. *p<0.05 v control group (time 0); †p<0.05 v IL-1β group (time 4 h).

Figure 3  .

Figure 3  

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Localisation of intercellular adhesion molecule (ICAM) 1 immunostaining in inflamed rat mesenteric postcapillary venules. Typical micrographs of the ileal section of rat mesentery whole mounts. (A) Preparations excised two hours post-administration of 20 ng rat interleukin (IL) 1β from an animal injected with control mouse IgG 10 minutes prior to sacrifice. (B) As for A, but the rat was injected with 2 mg/kg mouse antirat ICAM-1 10 minutes prior to sacrifice and tissue collection. Note the brown staining around the endothelium of the postcapillary venule. (C) As in B, but the rat was treated with 100 µg/kg subcutaneous dexamethasone one hour prior to intraperitoneal injection of IL-1β. Note the scarce brown immunostaining compared with B. Pictures are representative of five distinct preparations. L, vessel lumen. Bar, 30 µm.

Figure 4  .

Figure 4  

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Involvement of KC in interleukin (IL) 1β induced cell adhesion and emigration. (A) Animals were treated with 2 mg goat IgG or goat antirat rat KC polyclonal antibody one hour prior to IL-1β administration (20 ng intraperitoneally). Mesenteries were exposed two hours later, and the degree of cell adhesion (per 100 µm vessel wall) and emigration quantified by videomicroscopy. Data are mean (SEM); n=8 rats. *p<0.05 v mouse IgG group. (B) Detection of rat KC protein in the peritoneal cell free lavage fluids of control rats (n=4), rats injected with IL-1β (20 ng intraperitoneally; n=5), and animals pretreated with dexamethasone (DEX) one hour prior to IL-1β administration (n=5). Lavage fluids were collected two hours post-IL-1β. Data are mean (SEM). *p<0.05 v IL-1β group.

Figure 5  .

Figure 5  

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Localisation of KC immunostaining in inflamed rat mesenteric postcapillary venules. Typical micrographs of the ileal section of rat mesentery whole mounts. (A) Preparations excised two hours after administration of 20 ng rat interleukin (IL) 1β and incubated with non-specific rabbit IgG. Arrowheads indicate some of the perivenular mast cells. (B) As in A, but the tissue was incubated with specific rabbit antirat KC IgG. Arrowheads indicate some of the perivenular mast cells which stained for KC. (C) As in B, but the rat was treated with 100 µg/kg subcutaneous dexamethasone one hour prior to intraperitoneal injection of IL-1β. Arrowheads indicate some of the perivenular mast cells which appeared less stained for KC when compared with B. Pictures are representative of four distinct preparations. L, vessel lumen. Bar, 25 µm.

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