ESTABLISHMENT AND CHARACTERIZATION OF CELL-LINE LSV5 THAT RETAINS THEALTERED ADHESIVE PROPERTIES OF HUMAN JUNCTIONAL EPIDERMOLYSIS-BULLOSAKERATINOCYTES

Herlitz junctional epidermolysis bullosa (H-JEB) is characterized by h ampered expression of the adhesion ligand laminin-5. Thus far, analysi s of the processes underlying the epithelial-mesenchymal dysadhesion m arking this disease has been limited by the reduced growth and adhesiv e 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 enc odes the gamma 2 chain of laminin-5, Cell lines (LSV) derived from inf ected keratinocytes maintain a stable karyotype, grow independent of 3 T3 feeder layers and are not tumorigenic. Further analysis of clone LS V5 showed an increased secretion of laminin-6 and fibronectin compared to normal keratinocytes. Similar to parental H-JEB keratinocytes, the se cells regenerate stratified epidermis in vitro and, in in vivo mode ls, they synthesize a basement membrane lacking laminin-5. LSV cells s how hypermotility and reduced adhesive properties resulting from an in complete association with the underlying culture substrate. These resu lts demonstrate that LSV5 cells retain the pathologic phenotype of H-J EB keratinocytes and can serve as a model system to study the adhesion processes mediated by laminin-5. (C) 1996 Academic Press, Inc.