To evaluate the feasibility of using a replication-deficient recombinant adenovirus to transfer human genes to the human endothelium, human umbilical vein endothelial cells were infected in vitro with adenovirus vectors containing the lacZ gene or a human α1-antitrypsin (α1AT) cDNA. After in vitro infection with the lacZ adenovirus vector, cultured endothelial cells expressed β-galactosidase. In parallel studies with the α1AT adenovirus vector, infected cells expressed human α1AT transcripts, as evidenced by in situ hybridization and Northern analysis, and de novo synthesized and secreted glycosylated, functional α1AT within 6 hr of infection, as shown by [35S]methionine labeling and immunoprecipitation. Quantification of the culture supernatants demonstrated 0.3-0.6 μg of human α1AT secreted per 106 cells in 24 hr, for at least 14 days after adenovirus vector infection. To demonstrate the feasibility of direct transfer of genes into endothelial cells in human blood vessels, lacZ or α1AT adenovirus vectors were placed in the lumen of intact human umbilical veins ex vivo. Histologic evaluation of the veins after 24 hr demonstrated transfer and expression of the lacZ gene specifically to the endothelium. α1AT adenovirus infection resulted both in expression of α1AT transcripts in the endothelium and in de novo synthesis and secretion of α1AT. Quantification of α1AT in the vein perfusates showed average levels of 13 μg/ml after 24 hr. These observations strongly support the feasibility of in vivo human gene transfer to the endothelium mediated by replication-deficient adenovirus vectors.
|Number of pages||5|
|Journal||Proceedings of the National Academy of Sciences of the United States of America|
|Publication status||Published - 31 Jul 1992|
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