The stromal microenvironment plays a pivotal role in the development and persistence of chronic inflammation. Here, we tested the hypothesis that stromal fibroblasts can modulate the ability of endothelial cells (EC) to recruit leukocytes in a site specific manner. We compared the abilities of fibroblasts isolated from either the synovium or skin of patients with rheumatoid arthritis to modify the recruitment of peripheral blood lymphocytes (PBL) by EC, with or without exposure to inflammatory cytokines. EC and fibroblasts were cultured on opposite sides of 0.4µm-pore filters for 24h; a combination of tumour necrosis factor-alpha and interferon-gamma was added for a further 24h when desired. Filters were incorporated into a flow chamber to assess the adhesive properties of PBL. Co-culture of unstimulated EC with synovial, but not dermal fibroblasts, led to an increase in the capture of flowing PBL. This adhesion was inhibited by blockade of α4-integrin or CXCR4 on PBL, indicating that capture was supported by α4β1-VCAM-1 interaction and stabilised by activation through SDF-1α(CXCL12). Antibody neutralisation of IL-6 during co-culture effectively abolished the ability of EC to recruit PBL. Cytokine-stimulated EC supported high levels of PBL adhesion, through presentation of VCAM-1, E-selectin and chemokine(s) in this case acting through CXCR3. Interestingly, co-culture with dermal fibroblasts caused a marked reduction in cytokine-induced adhesion, whereas synovial fibroblasts induced a slight augmentation. In the dermal co-cultures, neutralisation of IL-6 caused a partial recovery of PBL adhesion, which was complete when both IL-6 and TGF-β were neutralised. We conclude that resting stromal fibroblasts can regulate the cytokine-sensitivity of vascular endothelium, while fibroblasts isolated from sites of chronic inflammation bypass this and develop a directly inflammatory phenotype. Interestingly, we detected dual roles for IL-6 in regulating ‘inflammation’ depending on the surrounding milieu. Its action was stimulatory in otherwise unstimulated co-cultures with synovial fibroblasts, but in dermal co-cultures treated with cytokines, IL-6 combined with TGF-β to desensitise the endothelial cells.