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Blood, Vol. 96 No. 2 (July 15), 2000:
pp. 554-559
The in vivo kinetics of tissue factor messenger RNA expression
during human endotoxemia: relationship with activation of coagulation
Rendrik F. Franco,
Evert de Jonge,
Pascale E. P. Dekkers,
Janneke J. Timmerman,
C. Arnold Spek,
Sander J. H. van Deventer,
Peter van Deursen,
Liesbeth van Kerkhoff,
Bob van Gemen,
Hugo ten Cate,
Tom van der Poll, and
Pieter H. Reitsma
From the Laboratory for Experimental Internal Medicine and
Department of Intensive Care, Academic Medical Center, University of
Amsterdam, and Organon Teknika, Boxtel, The Netherlands.
Triggering of the tissue factor (TF)-dependent coagulation pathway
is considered to underlie the generation of a procoagulant state during
endotoxemia. To determine the in vivo pattern of monocytic TF messenger
RNA (mRNA) expression during endotoxemia, 10 healthy volunteers were
injected with lipopolysaccharide (LPS, 4 ng/kg) and blood was collected
before and 0.5, 1, 2, 3, 4, 6, 8, and 24 hours after LPS
administration. Total blood RNA was isolated and amplified by NASBA
(nucleic acid sequence-based amplification), followed by quantitation
of TF mRNA by an electrochemiluminescence (ECL) assay. To compare the
pattern of coagulation activation with the kinetics of monocytic TF
mRNA expression, we measured plasma levels of markers of thrombin
generation, thrombin-antithrombin (TAT) complexes, and prothrombin
fragment 1 + 2 (F1 + 2). Baseline value (mean ± SEM) of the
number of TF mRNA molecules per monocytic cell was 0.08 ± 0.02. A
progressive and significant (P < .0001) increase in TF
expression was observed after LPS injection (+0.5 hour:
0.3 ± 0.1, +1 hour: 1.3 ± 0.9, +2 hours: 4.1 ± 0.9),
peaking at +3 hours (10 ± 1.9 TF mRNA molecules per monocyte). As
TF mRNA levels increased, thrombin generation was augmented. Peak
levels of TAT and F1 + 2 were reached later (at t +4 hours) than
those of TF mRNA. TF mRNA, TAT, and F1 + 2 levels returned to
baseline after 24 hours. In conclusion, we used a NASBA/ECL-based
technique to quantify TF mRNA in whole blood during human endotoxemia
and observed a 125-fold increase in TF mRNA levels. Our data
demonstrate a pivotal role for enhanced TF gene activity in the
activation of coagulation after LPS challenge.
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