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Stromal cells differentially regulate neutrophil and lymphocyte recruitment through the endothelium.

McGettrick HM, Buckley CD, Filer A, Rainger GE, Nash GB - Immunology (2010)

Bottom Line: Stromal fibroblasts modify the initial recruitment of leucocytes by endothelial cells (EC), but their effects on subsequent transendothelial migration remain unclear.Fibroblasts had little effect on tumour necrosis factor-α-induced transendothelial migration of neutrophils, but tended to increase the efficiency of migration away from the endothelium.To avoid contact, we cultured EC and fibroblasts on separate 3-μm pore filters one above the other.

View Article: PubMed Central - PubMed

Affiliation: Centre for Cardiovascular Sciences, School of Clinical and Experimental Medicine, Medical School, The University of Birmingham, Birmingham, UK. h.m.mcgettrick@bham.ac.uk

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Effect of fibroblasts on the adhesion and migration of T-cell subsets under static conditions. Endothelial cells cultured alone or with fibroblasts for 24 hr and were subsequently left untreated (a, c) or stimulated with tumour necrosis factor-α plus interferon-γ (TNF + IFN) (b, d). Adhesion (a, b) and migration (c, d) were assessed at 24 hr and expressed as a percentage of cells added for peripheral blood lymphocytes (PBL), CD4+ or CD8+ T cells. Data are mean ± SEM from three independent experiments. In (b), analysis of variance (anova) shows a significant effect of fibroblasts on T-cell adhesion, P < 0·01. In (c) and (d), anova shows a significant effect of fibroblasts on migration, P < 0·05. * = P < 0·05 and ** = P < 0·01 compared with None by Dunnett test.
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fig03: Effect of fibroblasts on the adhesion and migration of T-cell subsets under static conditions. Endothelial cells cultured alone or with fibroblasts for 24 hr and were subsequently left untreated (a, c) or stimulated with tumour necrosis factor-α plus interferon-γ (TNF + IFN) (b, d). Adhesion (a, b) and migration (c, d) were assessed at 24 hr and expressed as a percentage of cells added for peripheral blood lymphocytes (PBL), CD4+ or CD8+ T cells. Data are mean ± SEM from three independent experiments. In (b), analysis of variance (anova) shows a significant effect of fibroblasts on T-cell adhesion, P < 0·01. In (c) and (d), anova shows a significant effect of fibroblasts on migration, P < 0·05. * = P < 0·05 and ** = P < 0·01 compared with None by Dunnett test.

Mentions: We examined the ability of fibroblasts to modify the adhesion and migration of lymphocytes through EC with or without TNF-α + IFN-γ. Under static conditions, in the absence of cytokine treatment, PBL and CD4+ or CD8+ T-cell subsets (enumerated separately by flow cytometry) tended to be bound less efficiently to EC co-cultured with fibroblasts compared with EC cultured alone (Fig. 3a). Few lymphocytes migrated through the unstimulated cultures, but again, fewer migrated through co-cultures than endothelial monocultures (Fig. 3b). Adhesion and transmigration were increased and less variable after cytokine treatment. Under these conditions, dermal fibroblasts significantly inhibited adhesion compared with EC cultured alone, and co-culture with either type of fibroblast markedly reduced transmigration (Fig. 3). Similar suppression of migration by fibroblasts was observed when TNF-α alone was used as the stimulus (data not shown).


Stromal cells differentially regulate neutrophil and lymphocyte recruitment through the endothelium.

McGettrick HM, Buckley CD, Filer A, Rainger GE, Nash GB - Immunology (2010)

Effect of fibroblasts on the adhesion and migration of T-cell subsets under static conditions. Endothelial cells cultured alone or with fibroblasts for 24 hr and were subsequently left untreated (a, c) or stimulated with tumour necrosis factor-α plus interferon-γ (TNF + IFN) (b, d). Adhesion (a, b) and migration (c, d) were assessed at 24 hr and expressed as a percentage of cells added for peripheral blood lymphocytes (PBL), CD4+ or CD8+ T cells. Data are mean ± SEM from three independent experiments. In (b), analysis of variance (anova) shows a significant effect of fibroblasts on T-cell adhesion, P < 0·01. In (c) and (d), anova shows a significant effect of fibroblasts on migration, P < 0·05. * = P < 0·05 and ** = P < 0·01 compared with None by Dunnett test.
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Related In: Results  -  Collection

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fig03: Effect of fibroblasts on the adhesion and migration of T-cell subsets under static conditions. Endothelial cells cultured alone or with fibroblasts for 24 hr and were subsequently left untreated (a, c) or stimulated with tumour necrosis factor-α plus interferon-γ (TNF + IFN) (b, d). Adhesion (a, b) and migration (c, d) were assessed at 24 hr and expressed as a percentage of cells added for peripheral blood lymphocytes (PBL), CD4+ or CD8+ T cells. Data are mean ± SEM from three independent experiments. In (b), analysis of variance (anova) shows a significant effect of fibroblasts on T-cell adhesion, P < 0·01. In (c) and (d), anova shows a significant effect of fibroblasts on migration, P < 0·05. * = P < 0·05 and ** = P < 0·01 compared with None by Dunnett test.
Mentions: We examined the ability of fibroblasts to modify the adhesion and migration of lymphocytes through EC with or without TNF-α + IFN-γ. Under static conditions, in the absence of cytokine treatment, PBL and CD4+ or CD8+ T-cell subsets (enumerated separately by flow cytometry) tended to be bound less efficiently to EC co-cultured with fibroblasts compared with EC cultured alone (Fig. 3a). Few lymphocytes migrated through the unstimulated cultures, but again, fewer migrated through co-cultures than endothelial monocultures (Fig. 3b). Adhesion and transmigration were increased and less variable after cytokine treatment. Under these conditions, dermal fibroblasts significantly inhibited adhesion compared with EC cultured alone, and co-culture with either type of fibroblast markedly reduced transmigration (Fig. 3). Similar suppression of migration by fibroblasts was observed when TNF-α alone was used as the stimulus (data not shown).

Bottom Line: Stromal fibroblasts modify the initial recruitment of leucocytes by endothelial cells (EC), but their effects on subsequent transendothelial migration remain unclear.Fibroblasts had little effect on tumour necrosis factor-α-induced transendothelial migration of neutrophils, but tended to increase the efficiency of migration away from the endothelium.To avoid contact, we cultured EC and fibroblasts on separate 3-μm pore filters one above the other.

View Article: PubMed Central - PubMed

Affiliation: Centre for Cardiovascular Sciences, School of Clinical and Experimental Medicine, Medical School, The University of Birmingham, Birmingham, UK. h.m.mcgettrick@bham.ac.uk

Show MeSH
Related in: MedlinePlus