Abstract
Background: Sepsis is a major world-wide medical problem with high morbidity and mortality. Gram-negative bacteria are among the most important pathogens of sepsis and their LPS content is regarded to be important for the systemic inflammatory reaction. The CD14 /myeloid differentiation factor 2 (MD-2 )/TLR4 complex plays a major role in the immune response to LPS . The aim of this study was to compare the effects of inhibiting MD-2 and CD14 on ultra-pure LPS - versus whole E. coli bacteria-induced responses. Methods: Fresh human whole blood was incubated with upLPS or whole E. coli bacteria in the presence of MD-2 or CD14 neutralizing monoclonal antibodies, or their respective controls, and/or the specific complement-inhibitor compstatin. Cytokines were measured by a multiplex (n = 27) assay. NFκB activity was examined in cells transfected with CD14 , MD-2 and/or Toll-like receptors. Results: LPS -induced cytokine response was efficiently and equally abolished by MD-2 and CD14 neutralization. In contrast, the response induced by whole E. coli bacteria was only modestly reduced by MD-2 neutralization, whereas CD14 neutralization was more efficient. Combination with compstatin enhanced the effect of MD-2 neutralization slightly. When compstatin was combined with CD14 neutralization, however, the response was virtually abolished for all cytokines, including IL-17, which was only inhibited by this combination. The MD-2 -independent effect observed for CD14 could not be explained by TLR2 signaling. Conclusion: Inhibition of CD14 is more efficient than inhibition of MD-2 on whole E. coli -induced cytokine response, suggesting CD14 to be a better target for intervention in Gram-negative sepsis, in particular when combined with complement inhibition.
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Akashi-Takamura,S., Miyake,K. (2008). TLR accessory molecules. Curr. Opin. Immunol. 20, 420-425.
Annane,D., Bellissant,E., Cavaillon,J.M. (2005). Septic shock. Lancet 365, 63-78.
Antal-Szalmas,P. (2000). Evaluation of CD14 in host defence. Eur. J Clin. Invest 30, 167-179.
Baumann,C.L., Aspalter,I.M., Sharif,O., Pichlmair,A., Bluml,S., Grebien,F., Bruckner,M., Pasierbek,P., Aumayr,K., Planyavsky,M., Bennett,K.L., Colinge,J., Knapp,S., Superti-Furga,G. (2010). CD14 is a coreceptor of Toll-like receptors 7 and 9. J Exp. Med. 207, 2689-2701.
Brekke,O.L., Christiansen,D., Fure,H., Fung,M., Mollnes,T.E. (2007). The role of complement C3 opsonization, C5a receptor, and CD14 in E. coli-induced up-regulation of granulocyte and monocyte CD11b/CD18 (CR3), phagocytosis, and oxidative burst in human whole blood. J Leukoc. Biol. 81, 1404-1413.
Brekke,O.L., Christiansen,D., Fure,H., Pharo,A., Fung,M., Riesenfeld,J., Mollnes,T.E. (2008). Combined inhibition of complement and CD14 abolish E. coli-induced cytokine-, chemokine- and growth factor-synthesis in human whole blood. Mol. Immunol 45, 3804-3813.
Cai,S., Zemans,R.L., Young,S.K., Worthen,G.S., Jeyaseelan,S. (2008). MD-2-Dependent and -Independent Neutrophil Accumulation During Escherichia coli Pneumonia. Am. J Respir. Cell Mol. Biol. 40, 701-709.
Chow,J.C., Young,D.W., Golenbock,D.T., Christ,W.J., Gusovsky,F. (1999). Toll-like receptor-4 mediates lipopolysaccharide-induced signal transduction. J Biol. Chem. 274, 10689-10692.
Chun,K.H., Seong,S.Y. (2010). CD14 but not MD2 transmit signals from DAMP. Int. Immunopharmacol. 10, 98-106.
Erridge,C., Kennedy,S., Spickett,C.M., Webb,D.J. (2008). Oxidised phospholipid inhibition of toll-like receptor (TLR) signalling is restricted to TLR2 and TLR4 - roles for CD14, LPS-binding protein and MD2 as targets for specificity of inhibition. J Biol. Chem. 283, 24748-24759.
Flierl,M.A., Rittirsch,D., Gao,H., Hoesel,L.M., Nadeau,B.A., Day,D.E., Zetoune,F.S., Sarma,J.V., Huber-Lang,M.S., Ferrara,J.L., Ward,P.A. (2008). Adverse functions of IL-17A in experimental sepsis. FASEB J 22, 2198-2205.
Freitas,A., ves-Filho,J.C., Victoni,T., Secher,T., Lemos,H.P., Sonego,F., Cunha,F.Q., Ryffel,B. (2009). IL-17 receptor signaling is required to control polymicrobial sepsis. J Immunol 182, 7846-7854.
Janot,L., Secher,T., Torres,D., Maillet,I., Pfeilschifter,J., Quesniaux,V.F., Landmann,R., Ryffel,B., Erard,F. (2008). CD14 Works with Toll-Like Receptor 2 to Contribute to Recognition and Control of Listeria monocytogenes Infection. J Infect. Dis. 198, 115-124.
Katragadda,M., Magotti,P., Sfyroera,G., Lambris,J.D. (2006). Hydrophobic effect and hydrogen bonds account for the improved activity of a complement inhibitor, compstatin. J Med Chem. 49, 4616-4622.
Lappegard,K.T., Christiansen,D., Pharo,A., Thorgersen,E.B., Hellerud,B.C., Lindstad,J., Nielsen,E.W., Bergseth,G., Fadnes,D., Abrahamsen,T.G., Høyby,E.A., Schejbel,L., Garred,P., Harboe,M., Mollnes,T.E. (2009). Human genetic deficiencies reveal the roles of complement in the inflammatory network: lessons from nature. Proc Natl Acad Sci U S A 106, 15861-15866.
Martin,G.S., Mannino,D.M., Eaton,S., Moss,M. (2003). The epidemiology of sepsis in the United States from 1979 through 2000. N. Engl. J Med. 348, 1546-1554.
Miyake,K. (2006). Roles for accessory molecules in microbial recognition by Toll-like receptors. J. Endotoxin. Res. 12, 195-204.
Mollnes,T.E., Brekke,O.L., Fung,M., Fure,H., Christiansen,D., Bergseth,G., Videm,V., Lappegard,K.T., Kohl,J., Lambris,J.D. (2002). Essential role of the C5a receptor in E coli-induced oxidative burst and phagocytosis revealed by a novel lepirudin-based human whole blood model of inflammation. Blood 100, 1869-1877.
Nahra,R., Dellinger,R.P. (2008). Targeting the lipopolysaccharides: still a matter of debate? Curr. Opin. Anaesthesiol. 21, 98-104.
Nakata,T., Yasuda,M., Fujita,M., Kataoka,H., Kiura,K., Sano,H., Shibata,K. (2006). CD14 directly binds to triacylated lipopeptides and facilitates recognition of the lipopeptides by the receptor complex of Toll-like receptors 2 and 1 without binding to the complex. Cell Microbiol. 8, 1899-1909.
Nilsen,N.J., Deininger,S., Nonstad,U., Skjeldal,F., Husebye,H., Rodionov,D., von,A.S., Hartung,T., Lien,E., Bakke,O., Espevik,T. (2008). Cellular trafficking of lipoteichoic acid and Toll-like receptor 2 in relation to signaling; role of CD14 and CD36. J Leukoc. Biol. 84, 280-291.
Ohnishi,T., Muroi,M., Tanamoto,K. (2001). N-linked glycosylations at Asn(26) and Asn(114) of human MD-2 are required for toll-like receptor 4-mediated activation of NF-kappaB by lipopolysaccharide. J Immunol 167, 3354-3359.
Pugin,J., Heumann,I.D., Tomasz,A., Kravchenko,V.V., Akamatsu,Y., Nishijima,M., Glauser,M.P., Tobias,P.S., Ulevitch,R.J. (1994). CD14 is a pattern recognition receptor. Immunity. 1, 509-516.
Remick,D.G. (2007). Pathophysiology of sepsis. Am. J Pathol. 170, 1435-1444.
Shimazu,R., Akashi,S., Ogata,H., Nagai,Y., Fukudome,K., Miyake,K., Kimoto,M. (1999). MD-2, a molecule that confers lipopolysaccharide responsiveness on Toll-like receptor 4. J Exp. Med 189, 1777-1782.
Silasi-Mansat,R., Zhu,H., Popescu,N.I., Peer,G., Sfyroera,G., Magotti,P., Ivanciu,L., Lupu,C., Mollnes,T.E., Taylor,F.B., Kinasewitz,G., Lambris,J.D., Lupu,F. (2010). Complement inhibition decreases the procoagulant response and confers organ protection in a baboon model of E. coli sepsis. Blood 116, 1002-1010.
Thorgersen,E.B., Hellerud,B.C., Nielsen,E.W., Barratt-Due,A., Fure,H., Lindstad,J.K., Pharo,A., Fosse,E., Tonnessen,T.I., Johansen,H.T., Castellheim,A., Mollnes,T.E. (2009). CD14 inhibition efficiently attenuates early inflammatory and hemostatic responses in Escherichia coli sepsis in pigs. FASEB J 24, 712-722.
Tidswell,M., Tillis,W., Larosa,S.P., Lynn,M., Wittek,A.E., Kao,R., Wheeler,J., Gogate,J., Opal,S.M. (2010). Phase 2 trial of eritoran tetrasodium (E5564), a Toll-like receptor 4 antagonist, in patients with severe sepsis. Crit Care Med. 38, 306-308.
Xu,R., Wang,R., Han,G., Wang,J., Chen,G., Wang,L., Li,X., Guo,R., Shen,B., Li,Y. (2010). Complement C5a regulates Interleukine-17 by affecting the crosstalk between DCs and gammadelta T cells in CLP-induced sepsis. Eur. J Immunol 40, 1079-1088.
Acknowledgments
The authors thank Judith Krey Ludviksen and Anne Pharo for the excellent performing of cytokine and TCC measurements.The authors acknowledge support by The Odd Fellow Foundation (TEM) and US Public Health Service Grants GM062134, AI030040, AI072106, AI068730 (JDL).
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Christiansen, D., Brekke, O.L., Stenvik, J., Lambris, J.D., Espevik, T., Mollnes, T.E. (2012). Differential Effect of Inhibiting MD-2 and CD14 on LPS- Versus Whole E. coli Bacteria-Induced Cytokine Responses in Human Blood. In: Lambris, J., Hajishengallis, G. (eds) Current Topics in Innate Immunity II. Advances in Experimental Medicine and Biology, vol 946. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-0106-3_14
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DOI: https://doi.org/10.1007/978-1-4614-0106-3_14
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