on April 28, 2005
Published online before print April 28, 2005, doi: 10.1161/01.ATV.0000168413.29874.d7
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Submitted on February 24, 2005
Accepted on April 14, 2005
Intracellular and Surface Distribution of Monocyte Tissue Factor. Application to Intersubject Variability
Elena M. Egorina *;
sterud
From the Department of Biochemistry (E.M.E., M.A.S., G.B., J.O.O., B.O.), Institute of Medical Biology, University of Tromsø, Norway; the Department of Immunology and Transfusion Medicine (F.X.E.G.), University Hospital of Northern Norway, Tromsø; and the Department of Biochemistry (B.P.-S., K.G.M.), University of Vermont, Burlington.
* To whom correspondence should be addressed. E-mail: egorina{at}fagmed.uit.no.
Objective--The high and low responder phenomenon describes individual differences in lipopolysaccharide (LPS)-induced monocyte tissue factor (TF) activity. We characterized patterns of intracellular accumulation, externalization, and shedding of TF in response to LPS in mononuclear cells (MNCs) from high responders (HRs) and low responders (LRs).
Methods and Results--After 2 hours of LPS stimulation of whole blood, flow cytometry analyses revealed a larger population of TF-positive monocytes in HRs (32.0±3.5%) versus LRs (11.2±1.2%; P
0.05), along with a stronger mean fluorescence intensity of TF signal in HRs (7.1±0.5AU) compared with LRs (5.4±0.4AU; P0.05). The LPS-treated blood of the HR group contained 2-fold more TF-positive microparticles than LRs. In-cell Western assay demonstrated higher intracellular accumulation of TF in mononuclear cells (MNCs) from LRs because LPS induced a 3.7-fold increase of total TF levels in LRs versus a 1.5-fold increase in HRs. In contrast, in response to LPS stimulation, MNCs from HRs exhibited a 4-fold induction of surface TF, whereas MNCs from LRs only had a minor increase in surface TF levels.
Conclusions--The higher availability of surface TF antigen on MNCs from HRs and TF-containing microparticles might make these individuals more susceptible to hypercoagulation.
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