All Title Author
Keywords Abstract

PLOS ONE  2013 

Outer Membrane Vesicles Derived from Escherichia coli Up-Regulate Expression of Endothelial Cell Adhesion Molecules In Vitro and In Vivo

DOI: 10.1371/journal.pone.0059276

Full-Text   Cite this paper   Add to My Lib


Escherichia coli, as one of the gut microbiota, can evoke severe inflammatory diseases including peritonitis and sepsis. Gram-negative bacteria including E. coli constitutively release nano-sized outer membrane vesicles (OMVs). Although E. coli OMVs can induce the inflammatory responses without live bacteria, the effect of E. coli OMVs in vivo on endothelial cell function has not been previously elucidated. In this study, we show that bacteria-free OMVs increased the expression of endothelial intercellular adhesion molecule-1 (ICAM-1), E-selectin and vascular cell adhesion molecule-1, and enhanced the leukocyte binding on human microvascular endothelial cells in vitro. Inhibition of NF-κB and TLR4 reduced the expression of cell adhesion molecules in vitro. OMVs given intraperitoneally to the mice induced ICAM-1 expression and neutrophil sequestration in the lung endothelium, and the effects were reduced in ICAM-1-/- and TLR4-/- mice. When compared to free lipopolysaccharide, OMVs were more potent in inducing both ICAM-1 expression as well as leukocyte adhesion in vitro, and ICAM-1 expression and neutrophil sequestration in the lungs in vivo. This study shows that OMVs potently up-regulate functional cell adhesion molecules via NF-κB- and TLR4-dependent pathways, and that OMVs are more potent than free lipopolysaccharide.


[1]  Danese S, Dejana E, Fiocchi C (2007) Immune regulation by microvascular endothelial cells: directing innate and adaptive immunity, coagulation, and inflammation. J Immunol 178: 6017–6022.
[2]  Andonegui G, Zhou H, Bullard D, Kelly MM, Mullaly SC, et al. (2009) Mice that exclusively express TLR4 on endothelial cells can efficiently clear a lethal systemic Gram-negative bacterial infection. J Clin Invest 119: 1921–1930.
[3]  Harding M, Kubes P (2012) Innate immunity in the vasculature: interactions with pathogenic bacteria. Curr Opin Microbiol 15: 85–91.
[4]  Ley K, Laudanna C, Cybulsky MI, Nourshargh S (2007) Getting to the site of inflammation: the leukocyte adhesion cascade updated. Nat Rev Immunol 7: 678–689.
[5]  Rao RM, Yang L, Garcia-Cardena G, Luscinskas FW (2007) Endothelial-dependent mechanisms of leukocyte recruitment to the vascular wall. Circ Res 101: 234–247.
[6]  Cook-Mills JM, Deem TL (2005) Active participation of endothelial cells in inflammation. J Leukoc Biol 77: 487–495.
[7]  Aird WC (2003) The role of the endothelium in severe sepsis and multiple organ dysfunction syndrome. Blood 101: 3765–3777.
[8]  Davies MJ, Gordon JL, Gearing AJ, Pigott R, Woolf N, et al. (1993) The expression of the adhesion molecules ICAM-1, VCAM-1, PECAM, and E-selectin in human atherosclerosis. J Pathol 171: 223–229.
[9]  Haraldsen G, Kvale D, Lien B, Farstad IN, Brandtzaeg P (1996) Cytokine-regulated expression of E-selectin, intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) in human microvascular endothelial cells. J Immunol 156: 2558–2565.
[10]  Lee HM, Choi EJ, Kim JH, Kim TD, Kim YK, et al. (2010) A membranous form of ICAM-1 on exosomes efficiently blocks leukocyte adhesion to activated endothelial cells. Biochem Biophys Res Commun 397: 251–256.
[11]  Kaper JB, Nataro JP, Mobley HL (2004) Pathogenic Escherichia coli. Nat Rev Microbiol 2: 123–140.
[12]  May AK, Gleason TG, Sawyer RG, Pruett TL (2000) Contribution of Escherichia coli alpha-hemolysin to bacterial virulence and to intraperitoneal alterations in peritonitis. Infect Immun 68: 176–183.
[13]  Drake TA, Cheng J, Chang A, Taylor FB Jr (1993) Expression of tissue factor, thrombomodulin, and E-selectin in baboons with lethal Escherichia coli sepsis. Am J Pathol 142: 1458–1470.
[14]  Winzer K, Williams P (2003) Escherichia coli gets the message. Nat Med 9: 1118–1119.
[15]  Agace WW, Patarroyo M, Svensson M, Carlemalm E, Svanborg C (1995) Escherichia coli induces transuroepithelial neutrophil migration by an intercellular adhesion molecule-1-dependent mechanism. Infect Immun 63: 4054–4062.
[16]  Lee EY, Choi DS, Kim KP, Gho YS (2008) Proteomics in gram-negative bacterial outer membrane vesicles. Mass Spectrom Rev 27: 535–555.
[17]  Amano A, Takeuchi H, Furuta N (2010) Outer membrane vesicles function as offensive weapons in host-parasite interactions. Microbes Infect 12: 791–798.
[18]  Lee EY, Bang JY, Park GW, Choi DS, Kang JS, et al. (2007) Global proteomic profiling of native outer membrane vesicles derived from Escherichia coli. Proteomics 7: 3143–3153.
[19]  Choi DS, Kim DK, Choi SJ, Lee J, Choi JP, et al. (2011) Proteomic analysis of outer membrane vesicles derived from Pseudomonas aeruginosa. Proteomics 11: 3424–3429.
[20]  Bomberger JM, Maceachran DP, Coutermarsh BA, Ye S, O'Toole GA, et al. (2009) Long-distance delivery of bacterial virulence factors by Pseudomonas aeruginosa outer membrane vesicles. PLoS Pathog 5: e1000382.
[21]  Ellis TN, Leiman SA, Kuehn MJ (2010) Naturally produced outer membrane vesicles from Pseudomonas aeruginosa elicit a potent innate immune response via combined sensing of both lipopolysaccharide and protein components. Infect Immun 78: 3822–3831.
[22]  Kuehn MJ, Kesty NC (2005) Bacterial outer membrane vesicles and the host-pathogen interaction. Genes Dev 19: 2645–2655.
[23]  Ellis TN, Kuehn MJ (2010) Virulence and immunomodulatory roles of bacterial outer membrane vesicles. Microbiol Mol Biol Rev 74: 81–94.
[24]  Namork E, Brandtzaeg P (2002) Fatal meningococcal septicaemia with "blebbing" meningococcus. Lancet 360: 1741.
[25]  Mirlashari MR, Hagberg IA, Lyberg T (2002) Platelet-platelet and platelet-leukocyte interactions induced by outer membrane vesicles from N. meningitidis. Platelets 13: 91–99.
[26]  Mirlashari MR, Hoiby EA, Holst J, Lyberg T (2002) Outer membrane vesicles from Neisseria meningitidis. APMIS 110: 193–204.
[27]  Furuta N, Tsuda K, Omori H, Yoshimori T, Yoshimura F, et al. (2009) Porphyromonas gingivalis outer membrane vesicles enter human epithelial cells via an endocytic pathway and are sorted to lysosomal compartments. Infect Immun 77: 4187–4196.
[28]  Park KS, Choi KH, Kim YS, Hong BS, Kim OY, et al. (2010) Outer membrane vesicles derived from Escherichia coli induce systemic inflammatory response syndrome. PLoS One 5: e11334.
[29]  Parker H, Chitcholtan K, Hampton MB, Keenan JI (2010) Uptake of Helicobacter pylori outer membrane vesicles by gastric epithelial cells. Infect Immun 78: 5054–5061.
[30]  Kaparakis M, Turnbull L, Carneiro L, Firth S, Coleman HA, et al. (2010) Bacterial membrane vesicles deliver peptidoglycan to NOD1 in epithelial cells. Cell Microbiol 12: 372–385.
[31]  Srisatjaluk R, Doyle RJ, Justus DE (1999) Outer membrane vesicles of Porphyromonas gingivalis inhibit IFN-gamma-mediated MHC class II expression by human vascular endothelial cells. Microb Pathog 27: 81–91.
[32]  Flo TH, Halaas O, Lien E, Ryan L, Teti G, et al. (2000) Human toll-like receptor 2 mediates monocyte activation by Listeria monocytogenes, but not by group B streptococci or lipopolysaccharide. J Immunol 164: 2064–2069.
[33]  Somerville JE Jr, Cassiano L, Bainbridge B, Cunningham MD, Darveau RP (1996) A novel Escherichia coli lipid A mutant that produces an antiinflammatory lipopolysaccharide. J Clin Invest 97: 359–365.
[34]  Andonegui G, Bonder CS, Green F, Mullaly SC, Zbytnuik L, et al. (2003) Endothelium-derived Toll-like receptor-4 is the key molecule in LPS-induced neutrophil sequestration into lungs. J Clin Invest 111: 1011–1020.
[35]  Batra S, Cai S, Balamayooran G, Jeyaseelan S (2012) Intrapulmonary administration of leukotriene B(4) augments neutrophil accumulation and responses in the lung to Klebsiella infection in CXCL1 knockout mice. J Immunol 188: 3458–3468.
[36]  Xu H, Ye X, Steinberg H, Liu SF (2010) Selective blockade of endothelial NF-kappaB pathway differentially affects systemic inflammation and multiple organ dysfunction and injury in septic mice. J Pathol 220: 490–498.
[37]  Baumgarten G, Knuefermann P, Wrigge H, Putensen C, Stapel H, et al. (2006) Role of Toll-like receptor 4 for the pathogenesis of acute lung injury in Gram-negative sepsis. Eur J Anaesthesiol 23: 1041–1048.
[38]  Alves-Filho JC, de Freitas A, Russo M, Cunha FQ (2006) Toll-like receptor 4 signaling leads to neutrophil migration impairment in polymicrobial sepsis. Crit Care Med 34: 461–470.
[39]  Campbell JJ, Hedrick J, Zlotnik A, Siani MA, Thompson DA, et al. (1998) Chemokines and the arrest of lymphocytes rolling under flow conditions. Science 279: 381–384.
[40]  Hilbert DW, Pascal KE, Libby EK, Mordechai E, Adelson ME, et al. (2008) Uropathogenic Escherichia coli dominantly suppress the innate immune response of bladder epithelial cells by a lipopolysaccharide- and Toll-like receptor 4-independent pathway. Microbes Infect 10: 114–121.
[41]  Shin TS, Kim JH, Kim YS, Jeon SG, Zhu Z, et al. (2010) Extracellular vesicles are key intercellular mediators in the development of immune dysfunction to allergens in the airways. Allergy 65: 1256–1265.


comments powered by Disqus

Contact Us


微信:OALib Journal