title: Establishment and characterization of cell line LSV5 that retains the altered adhesive properties of human junctional epidermolysis bullosa keratinocytes.
abstract: Herlitz junctional epidermolysis bullosa (H-JEB) is characterized by hampered expression of the adhesion ligand laminin-5. Thus far, analysis of the processes underlying the epithelial-mesenchymal dysadhesion marking this disease has been limited by the reduced growth and adhesive capabilities of the epithelial cells derived from H-JEB patients. To overcome these difficulties, we used SV40 virus to immortalize H-JEB keratinocytes with a homozygous nonsense mutation in the gene that encodes the gamma2 chain of laminin-5. Cell lines (LSV) derived from infected keratinocytes maintain a stable karyotype, grow independent of 3T3 feeder layers and are not tumorigenic. Further analysis of clone LSV5 showed an increased secretion of laminin-6 and fibronectin compared to normal keratinocytes. Similar to parental H-JEB keratinocytes, these cells regenerate stratified epidermis in vitro and, in in vivo models, they synthesize a basement membrane lacking laminin-5. LSV cells show hypermotility and reduced adhesive properties resulting from an incomplete association with the underlying culture substrate. These results demonstrate that LSV5 cells retain the pathologic phenotype of H-JEB keratinocytes and can serve as a model system to study the adhesion processes mediated by laminin-5.
authors.affiliation: Faculté de Médecine, U385 INSERM, Nice, France.
authors.forename: C
authors.initials: C
authors.lastname: Miquel
authors.forename: L
authors.initials: L
authors.lastname: Gagnoux-Palacios
authors.forename: M
authors.initials: M
authors.lastname: Durand-Clement
authors.forename: P
authors.initials: P
authors.lastname: Marinkovich
authors.forename: J P
authors.initials: JP
authors.lastname: Ortonne
authors.forename: G
authors.initials: G
authors.lastname: Meneguzzi
mesh_terms: D016475:3T3 Cells / Q000166:cytology; D000818:Animals; D001485:Basement Membrane / Q000502:physiology; D002448:Cell Adhesion / Q000502:physiology; D015815:Cell Adhesion Molecules / Q000096:biosynthesis / Q000235:genetics; D002455:Cell Division / Q000502:physiology; D002461:Cell Line, Transformed / Q000166:cytology* / Q000648:ultrastructure; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D002465:Cell Movement / Q000502:physiology; D002472:Cell Transformation, Viral; D000078404:Epidermal Cells; D016109:Epidermolysis Bullosa, Junctional / Q000473:pathology*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007279:Injections, Subcutaneous; D007536:Isomerism; D015603:Keratinocytes / Q000166:cytology* / Q000648:ultrastructure; D007797:Laminin / Q000096:biosynthesis / Q000502:physiology; D051379:Mice; D008819:Mice, Nude; D008854:Microscopy, Electron; D009154:Mutation / Q000502:physiology; D010641:Phenotype; D013539:Simian virus 40 / Q000502:physiology; D000097794:Kalinin
chemical_list: D015815:Cell Adhesion Molecules; D007797:Laminin; D000097794:Kalinin