Polarity of secretion of alpha 1-antitrypsin by human respiratory epithelial cells after adenoviral transfer of a human alpha 1-antitrypsin cDNA. Academic Article uri icon

Overview

MeSH

  • Adenoviridae
  • Cell Polarity
  • Cells, Cultured
  • DNA, Complementary
  • Epithelial Cells
  • Gene Expression
  • Gene Transfer Techniques
  • Genetic Vectors
  • Glycosylation
  • Humans
  • In Vitro Techniques
  • Kidney
  • Protein Processing, Post-Translational
  • RNA, Messenger
  • Recombinant Proteins

MeSH Major

  • Epithelium
  • alpha 1-Antitrypsin

abstract

  • alpha 1-Antitrypsin (alpha AT) deficiency, a hereditary cause of progressive emphysema, can potentially be treated by transfer of a functional human alpha 1AT gene to the respiratory epithelium. For such an approach to be successful, alpha 1AT must be provided to both the interstitium and the epithelial surface--that is, the alpha 1AT directed by the transferred gene must be secreted to both the apical and basolateral surfaces of the epithelial cells. To evaluate this concept, a recombinant, replication-deficient adenoviral vector (Ad-alpha 1AT) containing a human alpha 1AT cDNA driven by an adenovirus major late promoter was used to infect Bet-1A human respiratory epithelial cells. The infected cells expressed Ad-alpha 1AT-directed mRNA transcripts and synthesized and secreted functional human alpha 1AT as shown by [35S]methionine labeling and immunoprecipitation of a 52-kD glycosylated human alpha 1AT molecule capable of interacting with neutrophil elastase, its natural substrate. Bet-1A cells grown on microporous polycarbonate membranes formed tight junctions (resistance > 150 omega x cm2). After infection with Ad-alpha 1AT, [35S]methionine labeling and enzyme-linked immunoassay demonstrated that alpha 1AT was secreted into both the apical and basolateral compartments, with an average apical to basolateral ratio of 1.9 +/- 0.2. Thus, human respiratory epithelial cells infected with a recombinant adenoviral vector containing a human alpha 1AT cDNA secrete alpha 1AT across both the apical and basolateral cell membranes, suggesting that the respiratory epithelium could serve as a target for in vivo gene therapy of alpha 1AT deficiency.

publication date

  • April 1995

has subject area

  • Adenoviridae
  • Cell Polarity
  • Cells, Cultured
  • DNA, Complementary
  • Epithelial Cells
  • Epithelium
  • Gene Expression
  • Gene Transfer Techniques
  • Genetic Vectors
  • Glycosylation
  • Humans
  • In Vitro Techniques
  • Kidney
  • Protein Processing, Post-Translational
  • RNA, Messenger
  • Recombinant Proteins
  • alpha 1-Antitrypsin

Research

keywords

  • Journal Article

Identity

Language

  • eng

Digital Object Identifier (DOI)

  • 10.1165/ajrcmb.12.4.7695917

PubMed ID

  • 7695917

Additional Document Info

start page

  • 379

end page

  • 384

volume

  • 12

number

  • 4