DEFICIENCY OF THE INTEGRIN BETA-4 SUBUNIT IN JUNCTIONAL EPIDERMOLYSIS-BULLOSA WITH PYLORIC ATRESIA - CONSEQUENCES FOR HEMIDESMOSOME FORMATION AND ADHESION PROPERTIES

Junctional epidermolysis bullosa (JEB) comprises a group of inherited autosomal recessive blistering disorders characterized by dermo-epider mal separation through the lamina lucida of the basement membrane, We identified a patient with JEB associated with pyloric atresia (PA), in whom the integrin beta 4 subunit was completely absent, At the ultras tructural level, the hemidesmosomes were reduced in number, appeared r udimentary and lacked a subbasal dense plate and frequently an inner a ttachment plaque, However, keratin filaments were still anchored to th e cytoplasmic plaque of the hemidesmosome, Immunofluorescence analysis showed that the beta 4 subunit was absent in the skin of the PA-JEB p atient, whereas the alpha 6 subunit appeared to be normally distribute d along the basement membrane zone, as were the other hemidesmosomal c omponents BP230, BP180 and HD1, Furthermore, the alpha 3 and beta 1 su bunits were not only detected at the lateral membranes of basal cells in PA-JEB skin, as in normal skin, but also along the basement membran e zone, The few hemidesmosome-like structures found in cultured kerati nocytes from the PA-JEB patient contained the hemidesmosomal component s BP230, BP180 and HD1, but not the integrin alpha 6 subunit, Like alp ha 3, this subunit was colocalized with vinculin in focal contacts at the ends of actin stress fibers. Immunoprecipitation analysis revealed that alpha 6 was associated with beta 1 on PA-JEB keratinocytes, wher eas normal human keratinocytes (NHKs) exclusively express alpha 6 beta 4 on their cell surface, The initial adhesion of PA-JEB and normal ke ratinocytes to laminin-1 and laminin-5, both ligands for alpha 6 beta 1 and alpha 6 beta 4, was similar, In migration assays, the PA-JEB ker atinocytes were more motile on laminin-5 than normal keratinocytes, Ou r observations indicate that the integrin alpha 6 beta 4 plays a cruci al role in the proper assembly of hemidesmosomes and in the stabilizat ion of the dermal-epidermal junction, The fragility of the skin and th e blistering in this patient appear to have been due to the deficiency of the integrin beta 4 submit, which results in the formation of too few and structurally abnormal hemidesmosomes.