Back to Search
Start Over
Recombinant Escherichia coli as a gene delivery vector into airway epithelial cells.
- Source :
-
Journal of controlled release : official journal of the Controlled Release Society [J Control Release] 2004 Jun 18; Vol. 97 (2), pp. 371-81. - Publication Year :
- 2004
-
Abstract
- To transfer genes into airway epithelial cells, we have generated auxotrophic dap Escherichia coli BM2710 mutant that expresses the invasin of Yersinia pseudotuberculosis and the listeriolysin of Listeria monocytogenes. E. coli BM2710 harboring a plasmid carrying the gfp gene was incubated with immortalized normal or cystic fibrosis (CF) airway epithelial cells or with primary bronchial epithelial cells grown as an explant-outgrowth cell culture model. Approximately 2% of immortalized cells expressed GFP. Few primary cells were transfected that were always poorly differentiated and located at the edge of the outgrowth. This was consistent with the expression of beta1-integrins only on these cells and with the required interaction for cell entry of E. coli expressing the invasin with beta1-integrins. The subsequent intracellular trafficking of E. coli BM2710 studied by confocal and electronic microscopy showed that the E. coli-containing phagosomes rapidly matured into phagolysosomes. This is the first demonstration that recombinant bacteria are able to transfer genes into primary airway epithelial cells, provided that they are able to invade the cells.<br /> (Copyright 2004 Elsevier B.V.)
- Subjects :
- Adhesins, Bacterial biosynthesis
Adhesins, Bacterial genetics
Bacterial Toxins biosynthesis
Bacterial Toxins genetics
Cells, Cultured
Cystic Fibrosis pathology
Genetic Vectors
Green Fluorescent Proteins biosynthesis
Green Fluorescent Proteins genetics
HeLa Cells
Heat-Shock Proteins biosynthesis
Heat-Shock Proteins genetics
Hemolysin Proteins
Humans
Integrin beta1 biosynthesis
Microscopy, Confocal
Microscopy, Electron
Mutation
Transfection
Bronchi cytology
Epithelial Cells metabolism
Escherichia coli genetics
Gene Transfer Techniques
Subjects
Details
- Language :
- English
- ISSN :
- 0168-3659
- Volume :
- 97
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Journal of controlled release : official journal of the Controlled Release Society
- Publication Type :
- Academic Journal
- Accession number :
- 15196763
- Full Text :
- https://doi.org/10.1016/j.jconrel.2004.03.025