Polarity of secretion of alpha 1-antitrypsin by human respiratory epithelial cells after adenoviral transfer of a human alpha 1-antitrypsin cDNA
| Autor: | W Siegfried, Michel Perricaudet, Melissa A. Rosenfeld, L D Stratford-Perricaudet, L Stier, Andrea Pavirani, J P Lecocq, Ronald G. Crystal |
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| Rok vydání: | 1995 |
| Předmět: |
Pulmonary and Respiratory Medicine
DNA Complementary Glycosylation Immunoprecipitation Clinical Biochemistry Genetic Vectors Alpha (ethology) Gene Expression Biology In Vitro Techniques Kidney Epithelium Viral vector Adenoviridae Cell polarity Humans Secretion RNA Messenger Molecular Biology Cells Cultured Tight junction Gene Transfer Techniques Cell Polarity Epithelial Cells Cell Biology Molecular biology Recombinant Proteins Neutrophil elastase alpha 1-Antitrypsin biology.protein Respiratory epithelium Protein Processing Post-Translational |
| Zdroj: | American journal of respiratory cell and molecular biology. 12(4) |
| ISSN: | 1044-1549 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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