Adenovirus-mediated transfer of a recombinant human alpha 1-antitrypsin cDNA to human endothelial cells. Academic Article uri icon

Overview

MeSH

  • Cells, Cultured
  • DNA
  • DNA, Recombinant
  • Gene Expression
  • Genetic Vectors
  • Humans
  • In Vitro Techniques
  • RNA, Messenger

MeSH Major

  • Adenoviruses, Human
  • Endothelium, Vascular
  • Transfection
  • alpha 1-Antitrypsin

abstract

  • 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 alpha 1-antitrypsin (alpha 1AT) cDNA. After in vitro infection with the lacZ adenovirus vector, cultured endothelial cells expressed beta-galactosidase. In parallel studies with the alpha 1AT adenovirus vector, infected cells expressed human alpha 1AT transcripts, as evidenced by in situ hybridization and Northern analysis, and de novo synthesized and secreted glycosylated, functional alpha 1AT within 6 hr of infection, as shown by [35S]methionine labeling and immunoprecipitation. Quantification of the culture supernatants demonstrated 0.3-0.6 micrograms of human alpha 1AT secreted per 10(6) 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 alpha 1AT 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. alpha 1AT adenovirus infection resulted both in expression of alpha 1AT transcripts in the endothelium and in de novo synthesis and secretion of alpha 1AT. Quantification of alpha 1AT in the vein perfusates showed average levels of 13 micrograms/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.

publication date

  • July 15, 1992

has subject area

  • Adenoviruses, Human
  • Cells, Cultured
  • DNA
  • DNA, Recombinant
  • Endothelium, Vascular
  • Gene Expression
  • Genetic Vectors
  • Humans
  • In Vitro Techniques
  • RNA, Messenger
  • Transfection
  • alpha 1-Antitrypsin

Research

keywords

  • Journal Article

Identity

Language

  • eng

PubMed Central ID

  • PMC49525

PubMed ID

  • 1631146

Additional Document Info

start page

  • 6482

end page

  • 6486

volume

  • 89

number

  • 14