Abstract
Lipid rafts are cholesterol- and sphingolipid-rich ordered microdomains distributed in the plasma membrane that participates in mammalian signal transduction pathways. To determine the role of lipid rafts in mediating interactions between enteropathogens and intestinal epithelial cells, membrane cholesterol was depleted from Caco-2 and IPEC-J2 cells using methyl-β-cyclodextrin. Cholesterol depletion significantly reduced Escherichia coli and Salmonella enteritidis adhesion and invasion into intestinal epithelial cells. Complementation with exogenous cholesterol restored bacterial adhesion to basal levels. We also evaluated the role of lipid rafts in the activation of Toll-like receptor 5 signaling by bacterial flagellin. Depleting membrane cholesterol reduced the ability of purified recombinant E. coli flagellin to activate TLR5 signaling in intestinal cells. These data suggest that both membrane cholesterol and lipid rafts play important roles in enteropathogen adhesion and contribute to the activation of innate immunity via flagellin–TLR5 signaling.
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Acknowledgments
This work was supported by grants from the Chinese National Science Foundation Grant (Nos. 31270171, 31072136 and 30771603), the Genetically Modified Organisms Technology Major Project of China (2014ZX08006-001B), the 948 programme Grant No. 2011-G24 from Ministry of Agriculture of the People’s Republic of China, a project founded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, the Program for ChangJiang Scholars and Innovative Research Team In University “PCSIRT” (IRT0978), a fund of excellent doctorial dissertations from Yangzhou University, a Program granted for Scientific Innovation Research of College Graduate in Jiangsu province (CXZZ13_0913).
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Communicated by Djamel Drider.
Mingxu Zhou and Qiangde Duan have contributed equally to this work.
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Zhou, M., Duan, Q., Li, Y. et al. Membrane cholesterol plays an important role in enteropathogen adhesion and the activation of innate immunity via flagellin–TLR5 signaling. Arch Microbiol 197, 797–803 (2015). https://doi.org/10.1007/s00203-015-1115-2
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DOI: https://doi.org/10.1007/s00203-015-1115-2