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Mediators Of Inflammation
Borzcka, K;Pciennikowska, A;Bjrkelund, H;Sobota, A;Kwiatkowska, K;
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- production stimulated with 1-1000ng/mL of smooth or rough LPS (sLPS or rLPS) and in sLPS binding to RAW264 and J744 cells. CD14 was indispensable for TNF- generation induced by a low concentration, 1ng/mL, of sLPS and rLPS. At higher doses of both LPS forms (100-1000ng/mL), TNF- release required CD14 to much lower extent. Among the two forms of LPS, rLPS-induced TNF- 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 1000ng/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- 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 production induced by high concentrations of sLPS and rLPS can be limited.