Induction of mesenchymal cell phenotypes in lung epithelial cells by adenovirus E1A.
Epithelial-mesenchymal transformation is now recognised as an important feature of tissue remodelling. The present report concerns the role of adenovirus infection in inducing this transformation in an animal model of chronic obstructive pulmonary disease. Guinea pig primary peripheral lung epithelial cells (PLECs) transfected with adenovirus E1A (E1A-PLECs) were compared to guinea pig normal lung fibroblasts (NLFs) transfected with E1A (E1A-NLFs). These cells were characterised by PCR, immunocytochemistry, electron microscopy, and Western and Northern blot analyses. Electrophoretic mobility shift assays were performed in order to examine nuclear factor (NF)-kappaB and activator protein (AP)-1 binding activities. E1A-PLECs and E1A-NLFs positive for E1A DNA, mRNA and protein expressed cytokeratin and vimentin but not smooth muscle alpha-actin. Both exhibited cuboidal morphology and junctional complexes, but did not contain lamellar bodies or express surfactant protein A, B or C mRNAs. These two cell types differed, however, in their NF-kappaB and AP-1 binding after lipopolysaccharide stimulation, possibly due to differences in the expression of the subunits that comprise these transcriptional complexes. E1A transfection results in the transformation of peripheral lung epithelial cells and normal lung fibroblasts to a phenotype intermediate between that of the two primary cells. It is postulated that this intermediate phenotype may play a major role in the remodelling of the airways in chronic obstructive pulmonary disease associated with persistence of adenovirus E1A DNA.
Behzad
A R
AR
James Hogg iCapture Center for Cardiovascular and Pulmonary Research, St Paul's Hospital, Vancouver, BC, Canada.
Morimoto
K
K
Gosselink
J
J
Green
J
J
Hogg
J C
JC
Hayashi
S
S
D000199:Actins / Q000378:metabolism; D017500:Adenovirus E1A Proteins / Q000502:physiology*; D000818:Animals; D001665:Binding Sites; D015152:Blotting, Northern; D015153:Blotting, Western; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D004279:DNA, Viral; D024202:Electrophoretic Mobility Shift Assay; D004847:Epithelial Cells / Q000378:metabolism / Q000821:virology; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D006168:Guinea Pigs; D007124:Immunoenzyme Techniques; D007633:Keratins / Q000378:metabolism; D008168:Lung / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D008648:Mesoderm / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D008854:Microscopy, Electron; D009130:Muscle, Smooth / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D016133:Polymerase Chain Reaction; D037662:Pulmonary Surfactant-Associated Protein A; D012333:RNA, Messenger; D012367:RNA, Viral; D018808:Transcription Factor AP-1; D014162:Transfection; D014746:Vimentin / Q000378:metabolism
D000199:Actins; D017500:Adenovirus E1A Proteins; D004279:DNA, Viral; D016328:NF-kappa B; D037662:Pulmonary Surfactant-Associated Protein A; D012333:RNA, Messenger; D012367:RNA, Viral; D018808:Transcription Factor AP-1; D014746:Vimentin; D007633:Keratins