Stromal cells differentially regulate neutrophil and lymphocyte recruitment through the endothelium.
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.
Affiliation: Centre for Cardiovascular Sciences, School of Clinical and Experimental Medicine, Medical School, The University of Birmingham, Birmingham, UK. email@example.comShow MeSH
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Mentions: We previously reported that adhesion of flowing lymphocytes to EC that had been treated with TNF-α + IFN-γ was mainly supported through α4β1-integrins binding to VCAM-1 on the EC, with a contribution from E-selectin; stabilization of this attachment required activation of lymphocytes through CXCR3 binding its ligands (CXCL9, CXCL10 and/or CXCL11).8 Using qPCR, we found that co-culture had no significant effect on mRNA levels for E-selectin, and tended to increase mRNA for VCAM-1 for EC treated with TNF-α + IFN-γ (Fig. 6a,b). As judged by flow cytometry, surface expression of E-selectin and VCAM-1 on the EC was not significantly different for cytokine-treated EC cultured alone or with fibroblasts (Fig. 6c,d). The RT-PCR for the CXCR3 ligands CXCL9, CXCL10 and CXCL11 showed their up-regulation in EC by the cytokines, but no difference in levels between EC cultured alone or with fibroblasts (data not shown). Hence, the functional loss observed in co-cultures could not be explained by changes in expression of endothelial adhesion receptors and chemokines identified as active in this model.
Affiliation: Centre for Cardiovascular Sciences, School of Clinical and Experimental Medicine, Medical School, The University of Birmingham, Birmingham, UK. firstname.lastname@example.org