CD14 Mediates Binding of High Doses of LPS but Is Dispensable for TNF-α Production

Activation of macrophages with lipopolysaccharide (LPS) involves a sequential engagement of serum LPS-binding protein (LBP), plasma membrane CD14, and TLR4/MD-2 signaling complex. We analyzed participation of CD14 in TNF-α production stimulated with 1–1000 ng/mL of smooth or rough LPS (sLPS or rLPS)...

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Main Authors: Kinga Borzęcka, Agnieszka Płóciennikowska, Hanna Björkelund, Andrzej Sobota, Katarzyna Kwiatkowska
Format: Article
Language:English
Published: Wiley 2013-01-01
Series:Mediators of Inflammation
Online Access:http://dx.doi.org/10.1155/2013/824919
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author Kinga Borzęcka
Agnieszka Płóciennikowska
Hanna Björkelund
Andrzej Sobota
Katarzyna Kwiatkowska
author_facet Kinga Borzęcka
Agnieszka Płóciennikowska
Hanna Björkelund
Andrzej Sobota
Katarzyna Kwiatkowska
author_sort Kinga Borzęcka
collection DOAJ
description Activation of macrophages with lipopolysaccharide (LPS) involves a sequential engagement of serum LPS-binding protein (LBP), plasma membrane CD14, and TLR4/MD-2 signaling complex. We analyzed participation of CD14 in TNF-α production stimulated with 1–1000 ng/mL of smooth or rough LPS (sLPS or rLPS) and in sLPS binding to RAW264 and J744 cells. CD14 was indispensable for TNF-α generation induced by a low concentration, 1 ng/mL, of sLPS and rLPS. At higher doses of both LPS forms (100–1000 ng/mL), TNF-α release required CD14 to much lower extent. Among the two forms of LPS, rLPS-induced TNF-α production was less CD14-dependent and could proceed in the absence of serum as an LBP source. On the other hand, the involvement of CD14 was crucial for the binding of 1000 ng/mL of sLPS judging from an inhibitory effect of the anti-CD14 antibody. The binding of sLPS was also strongly inhibited by dextran sulfate, a competitive ligand of scavenger receptors (SR). In the presence of dextran sulfate, sLPS-induced production of TNF-α was upregulated about 1.6-fold. The data indicate that CD14 together with SR participates in the binding of high doses of sLPS. However, CD14 contribution to TNF-α production induced by high concentrations of sLPS and rLPS can be limited.
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institution Kabale University
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spelling doaj-art-24c138c64e554cb4a3b9ca555176c8482025-08-20T03:54:56ZengWileyMediators of Inflammation0962-93511466-18612013-01-01201310.1155/2013/824919824919CD14 Mediates Binding of High Doses of LPS but Is Dispensable for TNF-α ProductionKinga Borzęcka0Agnieszka Płóciennikowska1Hanna Björkelund2Andrzej Sobota3Katarzyna Kwiatkowska4Department of Cell Biology, Nencki Institute of Experimental Biology, 3 Pasteur Street, 02-093 Warsaw, PolandDepartment of Cell Biology, Nencki Institute of Experimental Biology, 3 Pasteur Street, 02-093 Warsaw, PolandRidgeview Instruments AB, Skillsta 4, 740 20 Vänge, SwedenDepartment of Cell Biology, Nencki Institute of Experimental Biology, 3 Pasteur Street, 02-093 Warsaw, PolandDepartment of Cell Biology, Nencki Institute of Experimental Biology, 3 Pasteur Street, 02-093 Warsaw, PolandActivation of macrophages with lipopolysaccharide (LPS) involves a sequential engagement of serum LPS-binding protein (LBP), plasma membrane CD14, and TLR4/MD-2 signaling complex. We analyzed participation of CD14 in TNF-α production stimulated with 1–1000 ng/mL of smooth or rough LPS (sLPS or rLPS) and in sLPS binding to RAW264 and J744 cells. CD14 was indispensable for TNF-α generation induced by a low concentration, 1 ng/mL, of sLPS and rLPS. At higher doses of both LPS forms (100–1000 ng/mL), TNF-α release required CD14 to much lower extent. Among the two forms of LPS, rLPS-induced TNF-α production was less CD14-dependent and could proceed in the absence of serum as an LBP source. On the other hand, the involvement of CD14 was crucial for the binding of 1000 ng/mL of sLPS judging from an inhibitory effect of the anti-CD14 antibody. The binding of sLPS was also strongly inhibited by dextran sulfate, a competitive ligand of scavenger receptors (SR). In the presence of dextran sulfate, sLPS-induced production of TNF-α was upregulated about 1.6-fold. The data indicate that CD14 together with SR participates in the binding of high doses of sLPS. However, CD14 contribution to TNF-α production induced by high concentrations of sLPS and rLPS can be limited.http://dx.doi.org/10.1155/2013/824919
spellingShingle Kinga Borzęcka
Agnieszka Płóciennikowska
Hanna Björkelund
Andrzej Sobota
Katarzyna Kwiatkowska
CD14 Mediates Binding of High Doses of LPS but Is Dispensable for TNF-α Production
Mediators of Inflammation
title CD14 Mediates Binding of High Doses of LPS but Is Dispensable for TNF-α Production
title_full CD14 Mediates Binding of High Doses of LPS but Is Dispensable for TNF-α Production
title_fullStr CD14 Mediates Binding of High Doses of LPS but Is Dispensable for TNF-α Production
title_full_unstemmed CD14 Mediates Binding of High Doses of LPS but Is Dispensable for TNF-α Production
title_short CD14 Mediates Binding of High Doses of LPS but Is Dispensable for TNF-α Production
title_sort cd14 mediates binding of high doses of lps but is dispensable for tnf α production
url http://dx.doi.org/10.1155/2013/824919
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