Platelet factor 4 localization in carotid atherosclerotic plaques: correlation with clinical parameters

Stephanie Pitsilos; Jennifer Hunt; Emile R. Mohler; Anand M. Prabhakar; Mortimer Poncz; Jennine Dawicki; Tigran Z. Khalapyan; Megan L.Wolfe; Ronald Fairman; Marc Mitchell; Jeffrey Carpenter; Michael A. Golden; Douglas B. Cines; Bruce S. Sachais

doi:10.1160/TH03-02-0069

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2003; 90(06): 1112-1120
DOI: 10.1160/TH03-02-0069

Platelets and Blood Cells

Schattauer GmbH

Platelet factor 4 localization in carotid atherosclerotic plaques: correlation with clinical parameters

Authors

Stephanie Pitsilos

¹Departments of Pathology and Laboratory Medicine, Univ. of Pennsylvania, Philadelphia, USA
Jennifer Hunt

²Department of Pathology and Laboratory Medicine, University of Pittsburgh, Pittsburgh, USA
Emile R. Mohler

³Departments of Cardiology, Univ. of Pennsylvania, Philadelphia, USA
Anand M. Prabhakar

¹Departments of Pathology and Laboratory Medicine, Univ. of Pennsylvania, Philadelphia, USA
Mortimer Poncz

⁴Departments of Pediatrics, Univ. of Pennsylvania, Philadelphia, USA
Jennine Dawicki

¹Departments of Pathology and Laboratory Medicine, Univ. of Pennsylvania, Philadelphia, USA
Tigran Z. Khalapyan

³Departments of Cardiology, Univ. of Pennsylvania, Philadelphia, USA
Megan L.Wolfe

³Departments of Cardiology, Univ. of Pennsylvania, Philadelphia, USA
Ronald Fairman

⁵Departments of Surgery, Univ. of Pennsylvania, Philadelphia, USA
Marc Mitchell

⁵Departments of Surgery, Univ. of Pennsylvania, Philadelphia, USA
Jeffrey Carpenter

⁵Departments of Surgery, Univ. of Pennsylvania, Philadelphia, USA
Michael A. Golden

⁵Departments of Surgery, Univ. of Pennsylvania, Philadelphia, USA
Douglas B. Cines

¹Departments of Pathology and Laboratory Medicine, Univ. of Pennsylvania, Philadelphia, USA
Bruce S. Sachais

¹Departments of Pathology and Laboratory Medicine, Univ. of Pennsylvania, Philadelphia, USA

Abstract

Summary

Emerging evidence supports a role for platelets in the progression of atherosclerosis in addition to an involvement in thrombotic vascular occlusion. Platelet Factor 4 (PF4), a chemokine released by activated platelets, stimulates several pro-atherogenic processes. Therefore, we examined the localization of PF4 and the homologous protein, Neutrophil Activating Protein-2 (NAP-2) in lesions representing the evolution of human atherosclerotic plaques. Carotid plaques from 132 patients with critical carotid stenosis and 6 autopsy specimens were studied. Clinical, histologic and immunohistochemical data were analyzed using a χ²-test. PF4 was detected in the cytoplasm of luminal and neovascular endothelium, in macrophages and in regions of plaque calcification. The presence of PF4 in macrophages and neovascular endothelium correlated with lesion grade (p = 0.004; p = 0.044). Staining of macrophages for PF4 correlated with the presence of symptomatic atherosclerotic disease (p = 0.028). In early lesions, PF4 was commonly found in macrophages of early lesions (Grade I/II), whereas NAP-2 was rarely present.

In conclusion, correlation between PF4 deposition, lesion severity and symptomatic atherosclerosis suggests that persistent platelet activation may contribute to the evolution of athero-sclerotic vascular lesions. These studies support the rationale for the chronic use of anti-platelet therapy in patients at risk for developing symptomatic atherosclerosis.

Keywords

Atherosclerosis - platelet activation markers - vasculopathies - macrophage / phagocytosis

Full Text

References

References
1 Executive Summary of The Third Report of The National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, And Treatment of High Blood Cholesterol In Adults (Adult Treatment Panel III). Jama 2001; 285 (19) 2486-97.
2 Williams KJ, Tabas I. The response-to-retention hypothesis of early atherogenesis. Arterioscler Thromb Vasc Biol 1995; 15: 551-8.
3 Harker LA. Inhibitors of platelet function in the prevention of arterial thrombosis. Series Haematologica 1975; 8 (03) 105-24.
4 Sinzinger H, Kaliman J, Fitscha P, O’Grady J. Diminished platelet residence time on active human atherosclerotic lesions in-vivo—evidence for an optimal dose of aspirin?. Prostaglandins Leukotrienes & Essential Fatty Acids 1988; 34 (02) 89-93.
5 Chesterman CN, Berndt MC. Platelet and vessel wall interaction and the genesis of atherosclerosis. Clin Haematol 1986; 15 (02) 323-53.
6 Sevitt S. Platelets and foam cells in the evolution of atherosclerosis. Histological and immunohistological studies of human lesions. Atherosclerosis 1986; 61 (02) 107-15.
7 Sachais BS. Platelet-endothelial interactions in atherosclerosis. Curr Atheroscler Rep 2001; 3 (05) 412-6.
8 Holvoet P, Collen D. Thrombosis and atherosclerosis. Current Opinion In Lipidology 1997; 8 (05) 320-8.
9 Ross R. The pathogenesis of atherosclerosis: a perspective for the 1990s. Nature 1993; 362 6423 801-9.
10 Fuhrman B, Brook GJ, Aviram M. Activated platelets secrete a protein-like factor that stimulates oxidized-LDL receptor activity in macrophages. Journal of Lipid Research 1991; 32 (07) 1113-23.
11 Gawaz M, Neumann FJ, Dickfeld T. et al. Activated platelets induce monocyte chemo-tactic protein-1 secretion and surface expression of intercellular adhesion molecule-1 on endothelial cells. Circulation 1998; 98 (12) 1164-71.
12 Massberg S, Brand K, Gruner S. et al. A Critical Role of Platelet Adhesion in the Initiation of Atherosclerotic Lesion Formation. J Exp Med 2002; 196 (07) 887-96.
13 Sachais BS, Kuo A, Nassar T. et al. Platelet factor 4 binds to low-density lipoprotein receptors and disrupts the endocytic itinerary, resulting in retention of low-density lipo-protein on the cell surface. Blood 2002; 99: 3613-22.
14 Hunt JL, Fairman R, Mitchell ME. et al. Bone formation in carotid plaques: a clinicopatho-logical study. Stroke 2002; 33 (05) 1214-9.
15 Stary HC, Chandler AB, Dinsmore RE. et al. A definition of advanced types of atherosclerotic lesions and a histological classification of atherosclerosis. A report from the Committee on Vascular Lesions of the Council on Arteriosclerosis, American Heart Association. Arterioscler Thromb Vasc Biol 1995; 15 (09) 1512-31.
16 Arepally GM, Kamei S, Park KS. et al. Characterization of a murine monoclonal antibody that mimics heparin-induced thrombocytopenia antibodies. Blood 2000; 95 (05) 1533-40.
17 Rao AK, Niewiarowski S, James P. et al. Effect of heparin on the in vivo release and clearance of human platelet factor 4. Blood 1983; 61: 1208-14.
18 Busch C, Dawes J, Pepper DS, Wasteson A. Binding of platelet factor 4 to cultured human umbilical vein endothelial cells. Thrombosis Research 1980; 19 1-2 129-37.
19 Rucinski B, Niewiarowski S, Strzyzewski M, Holt JC, Mayo KH. Human platelet factor 4 and its C-terminal peptides: heparin binding and clearance from the circulation Thrombosis &amp. Haemostasis 1990; 63: 493-8.
20 Borgstrom P, Discipio R, Maione TE. Recombinant platelet factor 4, an angiogenic marker for human breast carcinoma. Anti-cancer Res 1998; 18 6A 4035-41.
21 Hansell P, Maione TE, Borgstrom P. Selective binding of platelet factor 4 to regions of active angiogenesis in vivo. Am J Physiol 1995; 269 3 Pt 2 H829-36.
22 Malek AM, Alper SL, Izumo S. Hemodynamic shear stress and its role in atherosclerosis. Jama 1999; 282 (21) 2035-42.
23 Wight TN. Cell biology of arterial proteogly-cans. Arteriosclerosis 1989; 9 (01) 1-20.
24 Deuel TF, Senior RM, Chang D. et al. Platelet factor 4 is chemotactic for neutrophils and monocytes. Proc Natl Acad Sci U S A 1981; 78 (07) 4584-7.
25 Nassar T, Sachais BS, Akkawi S. et al. Platelet Factor 4 Enhances the Binding of Oxidized Low-density Lipoprotein to Vascular Wall Cells. J Biol Chem 2003; 278 (08) 6187-93.
26 Collaborative overview of randomised trials of antiplatelet therapy—I: Prevention of death, myocardial infarction, and stroke by prolonged antiplatelet therapy in various categories of patients. Antiplatelet Trialists’ Collaboration. BMJ 1994; 308 6921 81-106.
27 Anonymous. Final report on the aspirin component of the ongoing Physicians’ Health Study. Steering Committee of the Physicians’ Health Study Research Group. N Engl J Med 1989; 321 (03) 129-35.