TY - JOUR T1 - Stimulation of NOD1 induces RIP2, TAK1 and p38 MAPK dependent pro-inflammatory signalling in human lung microvascular endothelial cells JF - European Respiratory Journal JO - Eur Respir J VL - 38 IS - Suppl 55 SP - 1687 AU - Timothy L. Gatheral AU - Laura Moreno AU - Daniel M. Reed AU - Rekha Badiger AU - Lucy K. Bailey AU - Neil Galloway-Phillipps AU - Jane A. Mitchell Y1 - 2011/09/01 UR - http://erj.ersjournals.com/content/38/Suppl_55/1687.abstract N2 - Gram-negative bacteria are an important cause of septic shock. NOD1 receptors recognise peptidoglycan in the bacterial cell wall and initiate pro-inflammatory responses. We have previously shown in rodents that stimulation of NOD1 induces vascular dysfunction in vitro and profound shock in vivo. In this study we investigated the role of NOD1 in human lung microvascular endothelial cells (HMVEC) which represent a site of key importance in the pathophysiology of sepsis and acute lung injury.HMVEC from healthy donors were cultured in 96-well plates. Cells were treated for 24 hours with vehicle ± LPS (TLR4) or iE-DAP (NOD1). In additional experiments cells were pre-treated for 1 hour with specific signalling inhibitors prior to addition of agonists (n=4-5). Cell activation was assessed by multiplex ELISA and by specific ELISAs for CXCL8 and 6-keto Prostaglandin F1α.iE-DAP induced significant release of CXCL8, 6-Keto Prostaglandin F1α, IL-1β, IL-2, and IFNγ. 5Z-7-oxo-zeaenol or BIRB0796 similarly inhibited responses to iE-DAP (Figure 1) or LPS (P<0.05;two-way ANOVA). By contrast, PP2 was more potent an inhibitor of iE-DAP (Figure 1) than LPS (p<0.05; two-way ANOVA).Figure 1In conclusion NOD1 is active in human microvascular endothelium and converges with TLR4 signalling at the level of TAK1 and p38 MAPK. NOD1 thus represents a potential target in the treatment of gram-negative sepsis. ER -