Effects of infection with recombinant adenovirus on human vascular endothelial and smooth muscle cells

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The plasminogen activation (PA) system is involved in vascular remodelling. Modulating its activity in vascular cells might be a way to interfere in processes such as angiogenesis and restenosis. Adenoviral vectors have become a favourable tool for direct gene transfer into vascular cells. In the interest of using adenoviral vectors to modulate plasminogen activator activity and endothelial and smooth muscle cell migration, we studied the effects of endothelial and smooth muscle cell transduction in vitro and in the umbilical vein ex vivo with a replication-defective adenoviral vector containing the β-galactosidase gene (AdCMVLacZ). Segments of the umbilical vein were infected with AdCMVLacZ (109-1010 pfu/ml). After 48 h strong β-galactosidase expression could be observed in the vessel wall, which was restricted to the endothelial layer. Although some heterogeneity in the transduction throughout the vessel could be seen, β-galactosidase expression was detectable for 21 days in explant. Infection of human vascular endothelial cells (HUVEC) with recombinant adenoviral vectors is a dose and time dependent process. To achieve 100% infection of cultured HUVEC after a 30 min infection period, adenovirus concentrations ranging from 1010 to 5.1010 pfu/ml are required. To achieve a similar infection of HUVSMC a concentration of 2.109 to 1010 is sufficient. Expression of β-galactosidase can be detected for at least 14 days. Effects of transduction of HUVEC with AdCMVLacZ on iferation, morphology and monolayer integrity were also studied. At high virus concentrations (> 1010 pfu/ml) an inhibitory effect on cell proliferation was detected and cell morphology displayed many giant cells, often polynuclear, in these cultures. Infection of confluent HUVEC monolayers had, apart from a rapid transient effect during the infection procedure, little effect on the permeability of these monolayers. Only when the monolayers were infected using ccncentrations of 1010 pfu/ml or more, an increase in permeability of these monolayers could be observed.