Salmonella typhimurium translocates flagellin across intestinal epithelia, inducing a proinflammatory response

@article{citeulike:1642416, abstract = {This study investigated whether soluble paracrine factors mediated Salmonella-induced IL-8 expression in polarized model intestinal epithelia. We found that the basolateral media of model epithelia that had been apically infected with Salmonella typhimurium for a short period (10 minutes) could activate IL-8 secretion in virgin model epithelia, demonstrating that a proinflammatory factor (PIF) was indeed present. Initial characterization found that PIF was a heat-stable protein with a molecular mass of about 50 kDa that acts on the basolateral, but not apical, surface of model intestinal epithelia to elicit IL-8 secretion. PIF was not present in the media of model epithelia stimulated with other inducers of IL-8 secretion (TNF-{alpha} or carbachol) but was present in S. typhimurium supernatants, indicating PIF is of bacterial origin. PIF was purified from bacterial culture supernatants by anion/cation exchange chromatography and SDS-PAGE and found by using microsequencing to be the protein flagellin. In support of this finding, flagellin-deficient S. typhimurium mutants did not secrete detectable levels of PIF (i.e., a bioactivity that induced IL-8 secretion when placed basolaterally on model epithelia). Furthermore, viable flagellin-deficient mutant organisms (fliC/fljB and flhD) failed to elicit IL-8 secretion when added apically to model intestinal epithelia. These findings indicate that translocation of flagellin across epithelia, subsequent to apical epithelial-S. typhimurium interaction, is likely a major means of activating a mucosal inflammatory response.}, author = {Gewirtz, Andrew T. and Simon, Peter O. and Schmitt, Clare K. and Taylor, Laura J. and Hagedorn, Curt H. and O'Brien, Alison D. and Neish, Andrew S. and Madara, James L. }, citeulike-article-id = {1642416}, journal = {J. Clin. Invest.}, keywords = {flagellin-biphasic}, month = {January}, number = {1}, pages = {99--109}, posted-at = {2007-09-10 16:30:29}, priority = {2}, title = {Salmonella typhimurium translocates flagellin across intestinal epithelia, inducing a proinflammatory response}, url = {http://www.jci.org/cgi/content/abstract/107/1/99}, volume = {107}, year = {2001} }

TY - JOUR ID - citeulike:1642416 L3 - citeulike-article-id:1642416 TI - Salmonella typhimurium translocates flagellin across intestinal epithelia, inducing a proinflammatory response JF - J. Clin. Invest. VL - 107 IS - 1 SP - 99 EP - 109 N2 - This study investigated whether soluble paracrine factors mediated Salmonella-induced IL-8 expression in polarized model intestinal epithelia. We found that the basolateral media of model epithelia that had been apically infected with Salmonella typhimurium for a short period (10 minutes) could activate IL-8 secretion in virgin model epithelia, demonstrating that a proinflammatory factor (PIF) was indeed present. Initial characterization found that PIF was a heat-stable protein with a molecular mass of about 50 kDa that acts on the basolateral, but not apical, surface of model intestinal epithelia to elicit IL-8 secretion. PIF was not present in the media of model epithelia stimulated with other inducers of IL-8 secretion (TNF-{alpha} or carbachol) but was present in S. typhimurium supernatants, indicating PIF is of bacterial origin. PIF was purified from bacterial culture supernatants by anion/cation exchange chromatography and SDS-PAGE and found by using microsequencing to be the protein flagellin. In support of this finding, flagellin-deficient S. typhimurium mutants did not secrete detectable levels of PIF (i.e., a bioactivity that induced IL-8 secretion when placed basolaterally on model epithelia). Furthermore, viable flagellin-deficient mutant organisms (fliC/fljB and flhD) failed to elicit IL-8 secretion when added apically to model intestinal epithelia. These findings indicate that translocation of flagellin across epithelia, subsequent to apical epithelial-S. typhimurium interaction, is likely a major means of activating a mucosal inflammatory response. KW - flagellin-biphasic AU - Gewirtz, Andrew AU - Simon, Peter AU - Schmitt, Clare AU - Taylor, Laura AU - Hagedorn, Curt AU - O'Brien, Alison AU - Neish, Andrew AU - Madara, James PY - 2001/01/01/ UR - http://www.jci.org/cgi/content/abstract/107/1/99 ER -