Expression of von Willebrand factor by human pulmonary endothelial cells in vivo.

BACKGROUND AND OBJECTIVES: Whether von Willebrand factor (vWf) is variably expressed by endothelial cells (EC) of the human vascular tree or not is not known. Studies on animals showed that the varying degrees of vWf expression in pulmonary vessels was a reflection of EC heterogeneity. Neither the influence of age or sex nor that of pathophysiological factors such as pulmonary hypertension (PH) on vWf expression has been systematically analysed up to now. However, such information is essential for the design and delivery of site-selective drugs and genes as well as for the analysis of EC culture systems. METHODS: The variable degrees of vWf expression in lung tissue specimens from 64 patients (age: 6 weeks to 86 years) with and without PH were studied immunohistochemically and analysed statistically. RESULTS: CD31-specific antibody was used as a control stain for EC. It produced equally strong staining reactions in all pulmonary EC. In contrast, vWf-specific antibody yielded negative or weakly positive staining reactions in capillary EC. The staining intensities increased hand in hand with vessel calibres. Besides, they increased statistically significantly with age and PH. Sex had no influence on vWf expression. CONCLUSION: These findings show that (1) vWf expression by pulmonary EC is heterogeneous and specific for individual types of vessels, (2) age and PH enhance vWf expression, (3) vWf expression is indicative of altered EC activation but not of EC defects, and (4) elevated plasma levels of vWf correlate positively with raised coagulatory activities in older patients and in patients with PH.

Expression of the endothelial markers PECAM-1, vWf, and CD34 in vivo and in vitro.

EC culture models are essential to study pathological alterations of endothelial cells (ECs) in pulmonary vascular diseases under standardized conditions. Nevertheless, little is known about the spectrum of alterations of vessel-specific endothelial phenotypes in monolayer cultures. For the comparative study of endothelial markers in vivo and in vitro we investigated immunohistochemically the expression of PECAM-1, vWf, and CD34 by pulmonary ECs in vivo and in stimulated/unstimulated human umbilical vein endothelial cells (HU-VEC) and human pulmonary microvascular endothelial cells (HPMEC). In vivo, vessel type-specific expression patterns were found for vWf and CD34, while PECAM-1 was homogeneously and strongly expressed. While all HUVEC showed a marked vWf staining, about two-thirds of HPMEC exhibited a strong and the rest a moderate vWf staining. In both in vitro models all ECs were clearly PECAM-1-positive. However, only about 20% of the HUVEC and HPMEC were CD34-positive. Our results demonstrate the reduced expression of vessel type-specific endothelial phenotypes by endothelial monolayer cultures, stressing the need to improve culture conditions as well as develop cocultures and three-dimensional culture models. Moreover, the need for endothelial markers specific for single microvascular type ECs becomes obvious in order to establish cultures consisting of only one microvascular ECs subpopulation.

Correlation of age with in vivo expression of endothelial markers.

The importance of endothelial senescence as a pathogenetic factor in age-related vascular alterations has almost exclusively been studied in vitro. However, the in vitro-findings have rarely been compared with histomorphological changes in aging human tissue or in age-related degenerative diseases. Therefore, we compared the expression of the endothelial marker CD34 and the procoagulant protein von Willebrand factor (vWf) in lung endothelium by conventional immunohistochemistry and confocal laser scan microscopy (CLSM), taking the age of the patients into consideration. The staining reactions were statistically analysed by covariance analysis. With age the endothelial staining intensity of CD34 increased in arteries and veins, but decreased in arterioles, capillaries and venules. For vWf, on the contrary, the endothelial staining intensity increased with age in all types of vessels. CLSM confirmed a mosaic staining pattern. This study demonstrates age-associated phenotypical alterations of CD34 and vWf. Whether the down-regulation of CD34 correlates with an age-associated reduction of the angiogenetic properties of EC or an age-related over-expression of vWf as a relevant cofactor for the raised coagulatory activity and the increase in thrombotic diseases resp coronary heart disease in older patients, remains subject to debate.