Human monocytes lacking the membrane-bound form of the bacterial lipopolysaccharide (LPS) receptor CD14 can mount an LPS-induced oxidative burst response mediated by a soluble form of CD14.
Monocytes and macrophages express a glycosyl phosphatidylinositol (GPI)-anchored lipopolysaccharide (LPS) receptor on the cell surface which enables them to detect minute amounts of LPS released from Gram-negative bacteria. A soluble form of CD14 is also found free in serum, though its physiological function is unknown. the interaction of LPS with CD14 on the monocyte surface leads to an activation of the cells which is manifested in the sudden release of reactive oxygen species, a process referred to as an oxidative burst. In patients suffering from the condition known as paroxysmal nocturnal haemoglobinuria (PNH), the synthesis of GPI anchors is blocked in haematopoietic cells which are therefore unable to express GPI-linked proteins on their surface. In severe cases, over 90% of monocytes lack membrane-bound CD14, though normal levels of the soluble form of the receptor-sCD14-are found in the serum. Despite this lack of membrane-bound CD14, monocytes from PNH patients can respond to low concentrations of LPS. Here we show that the LPS-induced oxidative burst of these PNH monocytes requires a component present in serum. The serum-dependent activation can be inhibited by monoclonal antibodies to CD14, can be removed from the serum by passage over a matrix to which an anti-CD14 antibody has been bound, and the depleted serum can be reconstituted by the addition of either purified natural or purified recombinant soluble CD14. We conclude that an LPS-dependent oxidative burst in PNH monocytes can be mediated by soluble CD14.
Schütt
C
C
Institut für Immunologie und Transfusionsmedizin, Klinikum der Universität Greifswald, Germany.
Schilling
T
T
Grunwald
U
U
Stelter
F
F
Witt
S
S
Krüger
C
C
Jack
R S
RS
D000209:Acute-Phase Proteins; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002352:Carrier Proteins / Q000378:metabolism; D002462:Cell Membrane / Q000276:immunology; D006224:Cricetinae; D017931:DNA Primers / Q000235:genetics; D006457:Hemoglobinuria, Paroxysmal / Q000276:immunology / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D018950:Lipopolysaccharide Receptors / Q000235:genetics / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008562:Membrane Glycoproteins; D008969:Molecular Sequence Data; D009000:Monocytes / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016897:Respiratory Burst; D012995:Solubility; D000099231:Lipopolysaccharide-Binding Protein
D000209:Acute-Phase Proteins; D002352:Carrier Proteins; D017931:DNA Primers; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D000099231:Lipopolysaccharide-Binding Protein