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Lipopolysaccharide activation of human monocytes mediated by CD14, results in a coordinated synthesis of tissue factor, TNF- and IL-6

Department of Clinical Chemistry, Ullevaal University Hospital, Oslo, Norway

Å.B. Westvik

Department of Clinical Chemistry, Ullevaal University Hospital, Oslo, Norway

R. Øvstebø

Department of Clinical Chemistry, Ullevaal University Hospital, Oslo, Norway

G-B. Joø

Department of Clinical Chemistry, Ullevaal University Hospital, Oslo, Norway

C. Okkenhaug

Department of Clinical Chemistry, Ullevaal University Hospital, Oslo, Norway

P. Kierulf

Department of Clinical Chemistry, Ullevaal University Hospital, Oslo, Norway

We have investigated the role of CD14 in lipopolysaccharide (LPS) stimulation of monocytes to tissue factor (TF) synthesis, tumor necrosis factor (TNF-) antigen and interleukin-6 (IL-6) antigen release. Elutriation-purified monocytes or peripheral blood mononuclear cells (PBMC) preincubated with a blocking monoclonal antibody against CD14, and monocytes with a reduced expression of CD14 (phosphatidyl inositol specific phospholipase C (PI-PLC) treated monocytes and PBMC isolated from a patient suffering from paroxysmal nocturnal hemoglobinuria, PNH), were stimulated with increasing LPS concentrations. TNF- antigen and IL-6 antigen were measured in the supernatants, and the cells harvested for measurement of procoagulant activity (PCA) as well as TF, TNF- and IL-6 mRNA. We conclude that LPS stimulation of monocytes to TF, TNF- and IL-6 synthesis as well as PCA is totally dependent on CD14 at clinically relevant LPS concentrations (1-50 ng/ml). Moreover, we demonstrate a close correlation between induction of PCA and release of TNF-a and IL-6 antigen indicating a common signalling pathway of induction of these mediators.

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