Oxidative stress enhances IL-8 and inhibits CCL20 production from intestinal epithelial cells in response to bacterial flagellin

Ivison SM, Wang C, Himmel ME, Sheridan J, Delano J, Mayer ML, Yao Y, Kifayet A, Steiner TS. Oxidative stress enhances IL-8 and inhibits CCL20 production from intestinal epithelial cells in response to bacterial flagellin. Am J Physiol Gastrointest Liver Physiol 299: G733-G741, 2010. First published July 1, 2010; doi: 10.1152/ajpgi.00089.2010.-Intestinal epithelial cells act as innate immune sentinels, as the first cells that encounter diarrheal pathogens. They use pattern recognition molecules such as the Toll-like receptors (TLRs) to identify molecular signals found on microbes but not host cells or food components. TLRs cannot generally distinguish the molecular signals on pathogenic bacteria from those found in commensals, yet under healthy conditions epithelial immune responses are kept in check. We hypothesized that, in the setting of tissue damage or stress, intestinal epithelial cells would upregulate their responses to TLR ligands to reflect the greater need for immediate protection against pathogens. We treated Caco-2 cells with the TLR5 agonist flagellin in the presence or absence of H2O2 and measured chemokine production and intracellular signaling pathways. H2O2 increased flagellin-induced IL-8 (CXCL8) production in a dose-dependent manner. This was associated with synergistic phosphorylation of p38 MAP kinase and with prolonged I-kappa B degradation and NF-kappa B activation. The H2O2-mediated potentiation of IL-8 production required the activity of p38, tyrosine kinases, phospholipase C gamma, and intracellular calcium, but not protein kinase C or protein kinase D. H2O2 prolonged and augmented NF-kappa B activation by flagellin. In contrast to IL-8, CCL20 (MIP3 alpha) production by flagellin was reduced by H2O2, and this effect was not calcium dependent. Oxidative stress biases intestinal epithelial responses to flagellin, leading to increased production of IL-8 and decreased production of CCL20. This suggests that epithelial cells are capable of sensing the extracellular environment and adjusting their antimicrobial responses accordingly.