Thrombosis Research 97 (2000) 369–374 BRIEF COMMUNICATION In Situ von Willebrand Factor Staining in Human Arteries and Veins Gregory T. Jones and Andre ´ van Rij Department of Surgery, Otago Medical School, Dunedin, New Zealand. (Received 22 March 1999 by Editor C.N. Chesterman; revised/accepted 23 June 1999) tunity for making detailed immunomorphological Key Words: Atherosclerosis; Endothelium; Human; Immu- nohistochemistry; Vascular injury; von Willebrand factor examinations of human endothelial cells as they ap- pear in situ, and has already been applied to the examination of growth factors [5,6], structural pro- V on Willebrand factor (vWF) is a factor VIII- teins [5], and cell surface adhesion molecules [7]. related multimeric glycoprotein essential to It has been suggested that vWF may be heteroge- platelet adhesion at sites of vascular injury neously expressed in different vascular beds, and [1]. Its production by endothelial cells has resulted may even be absent at some sites [8]. It has also in it being used as a specific marker for these cells been proposed that such heterogeneity may result [2,3]. Detailed immunolocalisation of endothelial in variable subendothelial matrix vWF levels, which antigens like vWF is hindered, however, by the thin in turn may lead to differential platelet binding activ- profile these cells present in conventional histologi- ity of the vessel wall under various clinical situations cal sections. Examination of cultured endothelial [9]. The possibility that human endothelium may cells, which are viewed en face, has shown that vWF regulate local subendothelial vWF deposition differ- is heterogeneously localised within the endoplasmic entially between various vascular beds is clearly reticulum and Weibel-Palade (WP) bodies of these of interest. cells and may also be associated with extracellular This study aims at determining if there is any matrix (ECM) proteins of the basal lamina [3]. heterogeneity in endothelial cell vWF immunostain- En face examination clearly represents the opti- ing of human vessels from a variety of vascular beds mal orientation for detailed in situ examination of and different pathophysiological states. endothelial cells. Although the en face “Ha ¨ utchen” preparation is a relatively standard technique for producing intact endothelial monolayers mounted 1. Materials and Methods on a microscope slide [4], it was not until recently that it has been successfully combined with immuno- Both arterial and venous specimens were obtained histochemistry [5]. This novel modification of an for examination in this study. Arterial specimens established technique provides the intriguing oppor- consisted of abdominal aortae from organ donor patients (n=5, ages 9, 29, 35, 60, and 70 years), as Abbreviations: vWF, von Willebrand factor; WP, Weibel-Palade; well as atherosclerotic carotid (n=3), femoral ECM, extracellular matrix; PBS, phosphate-buffered saline; BSA, (n=2), and tibial (n=2) arteries. Venous specimens bovine serum albumin; DAB, diaminobenzidine tetrahydrochlo- ride. included relatively normal long saphenous vein seg- Corresponding author: Dr. Greg Jones, Surgery Department, Uni- ments from limbs amputated for ischemic arterial versity of Otago, PO Box 913, Dunedin, New Zealand. Tel: +64- disease (n=2) or superfluous long saphenous seg- 3-474-0999; Fax: +64-3-474-7622; E-mail: greg.jones@stonebow. otago.ac.nz. ments from autologous bypass graft procedures 0049-3848/00 $–see front matter  2000 Elsevier Science Ltd. All rights reserved. PII S0049-3848(99)00166-8