J Inflamm Res. 2026 Oct 27;19:635143. doi: 10.2147/JIR.S635143. eCollection 2026.
ABSTRACT
BACKGROUND: Patients with immune-mediated inflammatory diseases (IMIDs), such as rheumatoid arthritis (RA), inflammatory bowel disease (IBD), and systemic lupus erythematosus (SLE), face a markedly increased risk of cardiovascular disease (CVD). This is mainly driven by chronic systemic inflammation, which in turn triggers endothelial activation and dysfunction.
OBJECTIVE: We aimed to establish a pathophysiologically relevant in vitro endothelial activation model using sub-nanogram concentrations of cytokines to mimic the low-grade systemic inflammatory milieu observed in active IMIDs.
METHODS: Human umbilical vein endothelial cells (HUVECs) were exposed to a combination of three key proinflammatory cytokines at circulating pathophysiological concentrations, representative of active clinical disease -interleukin-1 beta (IL-1β) at 20 pg/mL, tumor necrosis factor (TNF-α) at 50 pg/mL, and interferon-gamma (IFN-γ) at 100 pg/mL- referred to as the MIX. Each individual cytokine was also tested, and a high-dose IL-1β control (10 ng/mL) was included as a standard supra-physiological positive control. Surface marker expression was analyzed by flow cytometry, and cytokine/chemokine release was quantified by multiplex assay.
RESULTS: At 24 h, the MIX triggered a particular endothelial activation state. Notably, it significantly upregulated key immunological surface markers, including vascular cell adhesion molecule-1 (VCAM-1), and intercellular adhesion molecule-1 (ICAM-1), compared to the untreated control. Furthermore, the MIX significantly increased the release of key chemokines, such as IL-8, interferon-gamma inducible protein-10 (IP-10), and monocyte chemotactic protein-1 (MCP-1), displaying a distinct cooperative profile compared to individual low-dose cytokines.
CONCLUSION: Our study demonstrates that the combination of IL-1β, TNF-α, and IFN-γ at pathophysiologically relevant, sub-nanogram concentrations, can successfully model key features of IMID-associated endothelial activation by driving immune marker upregulation and secretory activity, opening new avenues towards the development of physiologically relevant in vitro system for future research into IMID-associated CVD.
PMID:42967185 | PMC:PMC13723236 | DOI:10.2147/JIR.S635143