Article | Published:

CD40L stabilizes arterial thrombi by a β3 integrin–dependent mechanism

Nature Medicine volume 8, pages 247252 (2002) | Download Citation

Subjects

Abstract

CD40L, a member of the tumor necrosis factor family of ligands, plays a major role in immune responses via its receptor, CD40. Recently, CD40L has been detected on the surfaces of activated platelets and shown to activate endothelium. Here we further addressed the function of platelet CD40L. We show that absence of CD40L affects the stability of arterial thrombi and delays arterial occlusion in vivo. Infusion of recombinant soluble (rs)CD40L restored normal thrombosis, whereas rsCD40L lacking the KGD integrin-recognition sequence did not. CD40-deficient mice exhibited normal thrombogenesis. rsCD40L specifically bound to purified integrin αIIbβ3 and to activated platelets in a β3-dependent manner and induced platelet spreading. In addition, rsCD40L promoted the aggregation of either human or mouse platelets under high shear rates. Thus, CD40L appears to be an αIIbβ3 ligand, a platelet agonist, and necessary for stability of arterial thrombi.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    & CD40-CD40 ligand. J. Leukoc. Biol. 67, 2–17 (2000).

  2. 2.

    et al. CD40 ligand on activated platelets triggers an inflammatory reaction of endothelial cells. Nature 391, 591–594 (1998).

  3. 3.

    , & CD40 ligand and the hyper-IgM syndrome. in Primary Immunodeficiency Diseases A Molecular and Genetic Approach (eds. Ochs, H.D., Smith, C.I.E. & Puck, J.M.) 233–249 (Oxford University Press, New York, 1999).

  4. 4.

    et al. Requirement for CD154 in the progression of atherosclerosis. Nature Med. 5, 1313–1316 (1999).

  5. 5.

    & The CD40/CD154 receptor/ligand dyad. Cell Mol. Life Sci. 58, 4–43 (2001).

  6. 6.

    , , , & Biological function of CD40 on human endothelial cells: costimulation with CD40 ligand and interleukin-4 selectively induces expression of vascular cell adhesion molecule-1 and P-selectin resulting in preferential adhesion of lymphocytes. Immunology 100, 441–448 (2000).

  7. 7.

    , & CD40L-CD40 interactions regulate endothelial cell surface tissue factor and thrombomodulin expression. J. Leukoc. Biol. 63, 373–379 (1998).

  8. 8.

    , , , & The inflammatory action of CD40 ligand (CD154) expressed on activated human platelets is temporally limited by coexpressed CD40. Blood 98, 1047–1054 (2001).

  9. 9.

    et al. Enhanced levels of soluble and membrane-bound CD40 ligand in patients with unstable angina. Possible reflection of T lymphocyte and platelet involvement in the pathogenesis of acute coronary syndromes. Circulation 100, 614–620 (1999).

  10. 10.

    , , , & Activation of monocyte/macrophage functions related to acute atheroma complication by ligation of CD40: induction of collagenase, stromelysin, and tissue factor. Circulation 96, 396–399 (1997).

  11. 11.

    , & Role of platelet P-selectin and CD40 ligand in the induction of monocytic tissue factor expression. Arterioscler. Thromb. Vasc. Biol. 20, 2322–2328 (2000).

  12. 12.

    et al. P-selectin induces the expression of tissue factor on monocytes. Proc. Natl. Acad. Sci. USA 91, 8767–8771 (1994).

  13. 13.

    , , , & Pro-coagulant state resulting from high levels of soluble P-selectin in blood. Proc. Natl. Acad. Sci. USA 97, 13835–13840 (2000).

  14. 14.

    et al. A soluble form of TRAP (CD40 ligand) is rapidly released after T cell activation. Eur. J. Immunol. 25, 1749–1754 (1995).

  15. 15.

    et al. Design of potent and specific integrin antagonists. Peptide antagonists with high specificity for glycoprotein IIb-IIIa. J. Biol. Chem. 268, 1066–1073 (1993).

  16. 16.

    , , , & Thromboembolic complications after treatment with monoclonal antibody against CD40 ligand. Nature Med. 6, 114 (2000).

  17. 17.

    et al. A mouse model of severe von Willebrand disease: defects in hemostasis and thrombosis. Proc. Natl. Acad. Sci. USA 95, 9524–9529 (1998).

  18. 18.

    et al. Persistence of platelet thrombus formation in arterioles of mice lacking both von Willebrand factor and fibrinogen. J. Clin. Invest. 106, 385–392 (2000).

  19. 19.

    & An integrin receptor on normal and thrombasthenic platelets that binds thrombospondin. Blood 74, 2022–2027 (1989).

  20. 20.

    et al. Long-term acceptance of skin and cardiac allografts after blocking CD40 and CD28 pathways. Nature 381, 434–438 (1996).

  21. 21.

    et al. Treatment with humanized monoclonal antibody against CD154 prevents acute renal allograft rejection in nonhuman primates. Nature Med. 5, 686–693 (1999).

  22. 22.

    , , , & Inhibition of CD40 signaling limits evolution of established atherosclerosis in mice. Proc. Natl. Acad. Sci. USA 97, 7458–7463 (2000).

  23. 23.

    et al. Both early and delayed anti-CD40L antibody treatment induces a stable plaque phenotype. Proc. Natl. Acad. Sci. USA 97, 7464–7469 (2000).

  24. 24.

    et al. Ligation of CD40 induces the expression of vascular endothelial growth factor by endothelial cells and monocytes and promotes angiogenesis in vivo. Blood 96, 3801–3808 (2000).

  25. 25.

    et al. Blood-borne tissue factor: another view of thrombosis. Proc. Natl. Acad. Sci. USA 96, 2311–2315 (1999).

  26. 26.

    & New perspectives in cell adhesion: RGD and integrins. Science 238, 491–497 (1987).

  27. 27.

    et al. Barbourin. A GP IIb-IIIa-specific integrin antagonist from the venom of Sistrurus m. barbouri. J. Biol. Chem. 266, 9359–9362 (1991).

  28. 28.

    , , , & Inhibition of fibrinogen binding to GP IIb-IIIa by a GP IIIa peptide. J. Biol. Chem. 266, 1415–1421 (1991).

  29. 29.

    et al. Ligands “activate” integrin α IIb β 3 (platelet GPIIb-IIIa). Cell 65, 409–416 (1991).

  30. 30.

    et al. The immune responses in CD40-deficient mice: impaired immunoglobulin class switching and germinal center formation. Immunity 1, 167–178 (1994).

  31. 31.

    et al. β3-integrin-deficient mice are a model for Glanzmann thrombasthenia showing placental defects and reduced survival. J. Clin. Invest. 103, 229–238 (1999).

  32. 32.

    et al. High-affinity interactions of tumor necrosis factor receptor-associated factors (TRAFs) and CD40 require TRAF trimerization and CD40 multimerization. Biochemistry 38, 10168–10177 (1999).

  33. 33.

    et al. Platelets adhere to and translocate on von Willebrand factor presented by endothelium in stimulated veins. Blood 96, 3322–3328 (2000).

  34. 34.

    et al. Optimal antagonism of GPIIb/IIIa favors platelet adhesion by inhibiting thrombus growth. An ex vivo capillary perfusion chamber study in the guinea pig. Arterioscler. Thromb. Vasc. Biol. 16, 56–63 (1996).

  35. 35.

    , , & Growth and stability of thrombi in flowing citrated blood: assessment of platelet-surface interactions with computer-assisted morphometry. Thromb. Haemost. 60, 392–398 (1988).

  36. 36.

    & Platelet membrane defects in Glanzmann's thrombasthenia. Evidence for decreased amounts of two major glycoproteins. J. Clin. Invest. 60, 535–545 (1977).

Download references

Acknowledgements

We thank L. Cowan for help with the preparation of the manuscript. Supported in part by National Heart, Lung and Blood Institute of the National Institutes of Health grants P01 HL56949 and R37 HL41002 (to D.D.W.).

Author information

Affiliations

  1. The Center for Blood Research, Harvard Medical School, Boston, Massachusetts, USA

    • Patrick André
    • , Cécile V. Denis
    • , Jessie M. Papalia
    •  & Denisa D. Wagner
  2. Department of Pathology, Harvard Medical School, Boston, Massachusetts, USA

    • Patrick André
    • , Cécile V. Denis
    •  & Denisa D. Wagner
  3. COR Therapeutics, South San Francisco, California, USA

    • K.S. Srinivasa Prasad
    • , Ming He
    •  & David R. Phillips
  4. Howard Hughes Medical Institute, Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, Masschusetts, USA

    • Richard O. Hynes

Authors

  1. Search for Patrick André in:

  2. Search for K.S. Srinivasa Prasad in:

  3. Search for Cécile V. Denis in:

  4. Search for Ming He in:

  5. Search for Jessie M. Papalia in:

  6. Search for Richard O. Hynes in:

  7. Search for David R. Phillips in:

  8. Search for Denisa D. Wagner in:

Competing interests

D.P., K.S.S.P. and M.H. are employees of COR Therapeutics. P.A. became an employee of COR Therapeutics after completion of study.

Corresponding author

Correspondence to Denisa D. Wagner.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nm0302-247

Further reading