1. Effects on Coagulation and Fibrinolysis Induced by Influenza in Mice With a Reduced Capacity to Generate Activated Protein C and a Deficiency in Plasminogen Activator Inhibitor Type 1
by Tymen T. Keller, Koen F. van der Sluijs, Martijn D. de Kruif, Victor E. A. Gerdes, Joost C. M. Meijers, Sandrine Florquin, Tom van der Poll, Eric C. M. van Gorp, Dees P. M. Brandjes, Harry R. Büller, and Marcel Levi
Circulation Research
Volume 99(11):
November 24, 2006
Copyright © American Heart Association, Inc. All rights reserved.

2. Figure 1. Influenza-induced changes in lungs.
Figure 1. Influenza-induced changes in lungs. Histopathological analysis of lungs of influenza-infected C57BL/6 mice. Lung sections were stained with H&E (A and B), Ly-6 for granulocytes (C and D), or fibrin (E and F). Compared with control mice (A, C, and E), influenza-infected mice (B, D, and F) showed inflammation in the lungs (B) and enhanced granulocyte influx (D) and fibrin deposition (F). Slides are representative of 6 mice, 4 days after influenza inoculation. Magnification, ×40 for H&E and fibrin and ×200 for Ly-6.
Tymen T. Keller et al. Circ Res. 2006;99:
Copyright © American Heart Association, Inc. All rights reserved.

3. Figure 2. Influenza-induced changes in lungs.
Figure 2. Influenza-induced changes in lungs. Analysis of granulocyte influx and fibrin formation in lungs of influenza-infected C57Bl/6 mice. Lung sections were stained with Ly-6 for granulocytes or fibrin. Mice inoculated with PBS (white bars) or influenza (black bars) and euthanized on days 0, 2, and 4 showed more granulocytes (A) and increased number of intravascular thrombi (B). Data are presented as mean±SE; n=6 for each group. *P<0.05 vs control, **P<0.01 vs control.
Tymen T. Keller et al. Circ Res. 2006;99:
Copyright © American Heart Association, Inc. All rights reserved.

4. Figure 3. Coagulation and fibrinolysis in influenza-infected mice.
Figure 3. Coagulation and fibrinolysis in influenza-infected mice. Plasma levels of TAT-c (A) and PAI-1 (B) in C57BL/6 mice inoculated with control buffer (PBS), low-dose influenza (Infl1), or high-dose influenza (Infl2). Mice were euthanized on days 0, 2, and 4. Data are presented as mean±SE. **P<0.01 low dose vs control or high dose vs low dose.
Tymen T. Keller et al. Circ Res. 2006;99:
Copyright © American Heart Association, Inc. All rights reserved.

5. Figure 4. Influenza load in TMpro/pro mice and PAI-1−/− mice.
Figure 4. Influenza load in TMpro/pro mice and PAI-1−/− mice. TMpro/pro mice (A) and PAI-1−/− mice (B) were inoculated with influenza at baseline. Viral outgrowth in the lungs of control mice (○) and transgenic mice (•) was determined 4 days after inoculation. Viral load is expressed as log10 viral RNA copies per microgram of RNA. There was no difference in viral load between transgenic mice and WT mice after 4 days.
Tymen T. Keller et al. Circ Res. 2006;99:
Copyright © American Heart Association, Inc. All rights reserved.

6. Figure 5. Thrombin generation and fibrin degradation increases in influenza-infected TMpro/pro mice.
Figure 5. Thrombin generation and fibrin degradation increases in influenza-infected TMpro/pro mice. TAT-c (A), PAI-1 (B), and D-dimer (C) levels in plasma. TMpro/pro (•) and TM WT (□) mice were inoculated with influenza and euthanized 0, 2, 4, 8, and 14 days after inoculation with influenza. Data are presented as mean±SE. A significant increase in TAT-c and D-dimer was demonstrated in TMpro/pro mice compared with TM WT mice. No difference in PAI-1 levels were seen. **P<0.01 vs WT.
Tymen T. Keller et al. Circ Res. 2006;99:
Copyright © American Heart Association, Inc. All rights reserved.

7. Figure 6. Treatment with LMWH reduce the prothrombotic state.
Figure 6. Treatment with LMWH reduce the prothrombotic state. Plasma levels of TAT-c (A), PAI-1 (B), and D-dimer (C) in influenza-inoculated C57Bl/6 mice treated with PBS (gray bars) or LMWH (black bars). A second control group of C57Bl/6 mice was inoculated with PBS and treated with PBS during the experiment (white bars). Mice were euthanized 4 days after inoculation. Influenza increased TAT-c, PAI-1, and D-dimer levels. In influenza-infected mice, treatment with LMWH reduced TAT-c levels compared with PBS. Data are presented as mean±SE. **P<0.01 influenza vs PBS or influenza+PBS vs influenza+LMWH.
Tymen T. Keller et al. Circ Res. 2006;99:
Copyright © American Heart Association, Inc. All rights reserved.

8. Figure 7. Fibrin degradation increases in influenza-infected PAI−/− mice.
Figure 7. Fibrin degradation increases in influenza-infected PAI−/− mice. TAT-c (A), PAI-1 (B), and D-dimer (C) levels in plasma. PAI-1−/− (•) and PAI-1 WT (□) mice were inoculated with influenza and euthanized 0, 2, 4, 8, and 14 days after inoculation with influenza. Data are presented as mean±SE. PAI-1 levels were undetectable in PAI-1−/− mice but remained increased in control mice until 14 days after infection. D-dimer levels were significantly increased in PAI-1−/− compared with PAI-1 WT mice. No differences in TAT-c levels were seen. **P<0.01 vs WT.
Tymen T. Keller et al. Circ Res. 2006;99:
Copyright © American Heart Association, Inc. All rights reserved.

9. Figure 8. Treatment with tranexamic acid decrease D-dimer levels.
Figure 8. Treatment with tranexamic acid decrease D-dimer levels. Plasma levels of TAT-c (A), PAI-1 (B), and D-dimer (C) in C57BL/6 mice inoculated with influenza and treated with PBS (gray bars) or tranexamic acid (black bars). Control mice were inoculated with PBS and treated with PBS during the experiment (white bars). Mice were euthanized 4 days after influenza inoculation. Influenza increased TAT-c, PAI-1, and D-dimer levels. In influenza-infected mice, treatment with tranexamic acid decreased D-dimer levels compared with PBS. Data are presented as mean±SE. **P<0.01 influenza vs PBS or influenza+PBS vs influenza+tranexamic acid.
Tymen T. Keller et al. Circ Res. 2006;99:
Copyright © American Heart Association, Inc. All rights reserved.