Limits...
Fibroblasts from different sites may promote or inhibit recruitment of flowing lymphocytes by endothelial cells.

McGettrick HM, Smith E, Filer A, Kissane S, Salmon M, Buckley CD, Rainger GE, Nash GB - Eur. J. Immunol. (2009)

Bottom Line: Interestingly, co-culture with dermal fibroblasts caused a marked reduction in cytokine-induced adhesion, while synovial fibroblasts had variable effects depending on their source.Exogenous IL-6 was also found to inhibit response to TNF-alpha+IFN-gamma.Normal stromal fibroblasts appear to regulate the cytokine-sensitivity of vascular endothelium, while fibroblasts associated with chronic inflammation bypass this and develop a directly inflammatory phenotype.

View Article: PubMed Central - PubMed

Affiliation: Centre for Cardiovascular Sciences, The Medical School, The University of Birmingham, Birmingham, UK.

ABSTRACT
We examined the hypothesis that stromal fibroblasts modulate the ability of endothelial cells (EC) to recruit lymphocytes in a site-specific manner. PBL were perfused over HUVEC that had been cultured with fibroblasts isolated from the inflamed synovium or the skin of patients with rheumatoid arthritis or osteoarthritis, or from normal synovium, with or without exposure to the inflammatory cytokines TNF-alpha+IFN-gamma. Fibroblasts from inflamed synovium, but no others, caused unstimulated HUVEC to bind flowing lymphocytes. This adhesion was supported by alpha(4)beta(1)-VCAM-1 interaction and stabilised by activation of PBL through CXCR4-CXCL12. Antibody neutralisation of IL-6 during co-culture effectively abolished the ability of EC to bind lymphocytes. Cytokine-stimulated EC supported high levels of lymphocyte adhesion, through the presentation of VCAM-1, E-selectin and chemokine(s) acting through CXCR3. Interestingly, co-culture with dermal fibroblasts caused a marked reduction in cytokine-induced adhesion, while synovial fibroblasts had variable effects depending on their source. In the dermal co-cultures, neutralisation of IL-6 or TGF-beta caused partial recovery of cytokine-induced lymphocyte adhesion; this was complete when both were neutralised. Exogenous IL-6 was also found to inhibit response to TNF-alpha+IFN-gamma. Normal stromal fibroblasts appear to regulate the cytokine-sensitivity of vascular endothelium, while fibroblasts associated with chronic inflammation bypass this and develop a directly inflammatory phenotype. Actions of IL-6 might be pro-inflammatory or anti-inflammatory, depending on the local milieu.

Show MeSH

Related in: MedlinePlus

Effect of RA co-culture on the surface expression of E-selectin or VCAM-1 by EC. EC from cytokine treated co-cultures were stained with antibodies against (A) E-selectin or (B) VCAM-1 and surface expression was assessed by flow cytometry (expressed as MFI). Data are mean±SEM from three independent experiments, using three different donors for each cell type.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2821685&req=5

fig04: Effect of RA co-culture on the surface expression of E-selectin or VCAM-1 by EC. EC from cytokine treated co-cultures were stained with antibodies against (A) E-selectin or (B) VCAM-1 and surface expression was assessed by flow cytometry (expressed as MFI). Data are mean±SEM from three independent experiments, using three different donors for each cell type.

Mentions: Judged by flow cytometry, surface levels of E-selectin and VCAM-1 were barely above background for unstimulated EC co-cultured with either fibroblast type (data not shown). Cytokine-treated EC showed strong expression of these receptors, and there was a tendency towards a reduction in E-selectin and VCAM-1 surface expression on co-cultures compared with EC cultured alone (Fig. 4). However, these effects were not statistically significant and similar trends were observed for each type of fibroblast from RA donors.


Fibroblasts from different sites may promote or inhibit recruitment of flowing lymphocytes by endothelial cells.

McGettrick HM, Smith E, Filer A, Kissane S, Salmon M, Buckley CD, Rainger GE, Nash GB - Eur. J. Immunol. (2009)

Effect of RA co-culture on the surface expression of E-selectin or VCAM-1 by EC. EC from cytokine treated co-cultures were stained with antibodies against (A) E-selectin or (B) VCAM-1 and surface expression was assessed by flow cytometry (expressed as MFI). Data are mean±SEM from three independent experiments, using three different donors for each cell type.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2821685&req=5

fig04: Effect of RA co-culture on the surface expression of E-selectin or VCAM-1 by EC. EC from cytokine treated co-cultures were stained with antibodies against (A) E-selectin or (B) VCAM-1 and surface expression was assessed by flow cytometry (expressed as MFI). Data are mean±SEM from three independent experiments, using three different donors for each cell type.
Mentions: Judged by flow cytometry, surface levels of E-selectin and VCAM-1 were barely above background for unstimulated EC co-cultured with either fibroblast type (data not shown). Cytokine-treated EC showed strong expression of these receptors, and there was a tendency towards a reduction in E-selectin and VCAM-1 surface expression on co-cultures compared with EC cultured alone (Fig. 4). However, these effects were not statistically significant and similar trends were observed for each type of fibroblast from RA donors.

Bottom Line: Interestingly, co-culture with dermal fibroblasts caused a marked reduction in cytokine-induced adhesion, while synovial fibroblasts had variable effects depending on their source.Exogenous IL-6 was also found to inhibit response to TNF-alpha+IFN-gamma.Normal stromal fibroblasts appear to regulate the cytokine-sensitivity of vascular endothelium, while fibroblasts associated with chronic inflammation bypass this and develop a directly inflammatory phenotype.

View Article: PubMed Central - PubMed

Affiliation: Centre for Cardiovascular Sciences, The Medical School, The University of Birmingham, Birmingham, UK.

ABSTRACT
We examined the hypothesis that stromal fibroblasts modulate the ability of endothelial cells (EC) to recruit lymphocytes in a site-specific manner. PBL were perfused over HUVEC that had been cultured with fibroblasts isolated from the inflamed synovium or the skin of patients with rheumatoid arthritis or osteoarthritis, or from normal synovium, with or without exposure to the inflammatory cytokines TNF-alpha+IFN-gamma. Fibroblasts from inflamed synovium, but no others, caused unstimulated HUVEC to bind flowing lymphocytes. This adhesion was supported by alpha(4)beta(1)-VCAM-1 interaction and stabilised by activation of PBL through CXCR4-CXCL12. Antibody neutralisation of IL-6 during co-culture effectively abolished the ability of EC to bind lymphocytes. Cytokine-stimulated EC supported high levels of lymphocyte adhesion, through the presentation of VCAM-1, E-selectin and chemokine(s) acting through CXCR3. Interestingly, co-culture with dermal fibroblasts caused a marked reduction in cytokine-induced adhesion, while synovial fibroblasts had variable effects depending on their source. In the dermal co-cultures, neutralisation of IL-6 or TGF-beta caused partial recovery of cytokine-induced lymphocyte adhesion; this was complete when both were neutralised. Exogenous IL-6 was also found to inhibit response to TNF-alpha+IFN-gamma. Normal stromal fibroblasts appear to regulate the cytokine-sensitivity of vascular endothelium, while fibroblasts associated with chronic inflammation bypass this and develop a directly inflammatory phenotype. Actions of IL-6 might be pro-inflammatory or anti-inflammatory, depending on the local milieu.

Show MeSH
Related in: MedlinePlus