A MULTIPLEX ASSAY OF 26 CYTOKINES SECRETED BY BLOOD CELLS OF PATIENTS WITH LIVER CIRRHOSIS

Abstract

Cytokines play an important role in the pathogenesis of liver cirrhosis (LC), determining the disease severity as well as hepatic and extrahepatic complications. The present study aims to characterize the cytokine profile of blood cells in 20 LC patients with regard to the stage (class A on Child-Pugh, n = 13, and the class B+C, n = 7) and the etiology of the disease (viral LC, n = 12 and the LC of non-viral etiology, n = 8) using cytokine 26-plex assay. The control group consisted of 10 healthy donors. Cytokine concentrations were tested both in spontaneous and bacterial endotoxin (LPS) stimulated 24 hour blood cultures. Four functional groups: pro-/anti-inflammatory cytokines (IL-1β, TNFα, IL-1ra, IL-10), immunoregulatory cytokines (IL-2, IFNγ, IL-12, IL-4, IL-5, IL-6, IL-9, IL-13, IL-15, IL-17), growth factors (G-CSF, IL-7, FGF-β, PDGF, VEGF) and chemokines (IL-8, IP-10, MCP-1, MIP-1α, MIP-1β, RANTES, Eotaxin) involved in the pathogenesis of inflammation, have been analyzed. The blood cells of LC patients were shown to be active producers of cytokines and were characterized by increased cytokine levels from all four functional groups. The enhancement of many cytokines was associated with the disease severity and viral etiology. The increased cytokine production in both intact and LPS-stimulated cultures indicated preserved reactivity of blood cells to endotoxin. Overall, the data obtained suggest the contribution of circulating blood cells in supporting of inflammatory response and the potential diagnostic/prognostic significance of blood cell-derived cytokines in patients with LC.

Keywords

• multiplex assay
• cytokines
• chemokines
• growth factors
• blood cells
• liver cirrhosis