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

W. Siegfried, M. Rosenfeld, L. Stier, L. Stratford-Perricaudet, M. Perricaudet, A. Pavirani, J. P. Lecocq, Ronald Crystal

Research output: Contribution to journalArticle

22 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)379-384
Number of pages6
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume12
Issue number4
DOIs
Publication statusPublished - 1 Jan 1995
Externally publishedYes

Fingerprint

alpha 1-Antitrypsin
polycarbonate
Complementary DNA
Epithelial Cells
Methionine
Labeling
Genes
Gene therapy
Leukocyte Elastase
Respiratory Mucosa
Cell membranes
Cells
Membranes
Messenger RNA
Molecules
alpha 1-Antitrypsin Deficiency
Substrates
Enzymes
Tight Junctions
Emphysema

ASJC Scopus subject areas

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry
  • Cell Biology

Cite this

Polarity of secretion of alpha 1-antitrypsin by human respiratory epithelial cells after adenoviral transfer of a human alpha 1-antitrypsin cDNA. / Siegfried, W.; Rosenfeld, M.; Stier, L.; Stratford-Perricaudet, L.; Perricaudet, M.; Pavirani, A.; Lecocq, J. P.; Crystal, Ronald.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 12, No. 4, 01.01.1995, p. 379-384.

Research output: Contribution to journalArticle

Siegfried, W. ; Rosenfeld, M. ; Stier, L. ; Stratford-Perricaudet, L. ; Perricaudet, M. ; Pavirani, A. ; Lecocq, J. P. ; Crystal, Ronald. / Polarity of secretion of alpha 1-antitrypsin by human respiratory epithelial cells after adenoviral transfer of a human alpha 1-antitrypsin cDNA. In: American Journal of Respiratory Cell and Molecular Biology. 1995 ; Vol. 12, No. 4. pp. 379-384.
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