Pemphigoid, pemphigus and Pr antigens in adult human keratinocytes grown in nonviable substrates

Abstract

Isolated adult human keratinocytes were grown either on plastic coverslips or a nonviable basement membrane surface containing intact laminin, type IV and V collagens, and heparan sulfate proteoglycan and examined by indirect immunofluorescence for the expression of bullous pemphigoid, pemphigus and Prlh antigens. Initial cell suspensions had a mean of 23% and 30%, respectively of bullous pemphigoid and Prlh positive staining cells, while those stained with pemphigus serum were usually negative (19 of 22 series). Pemphigus antigen expressed as intercellular staining between keratinocytes within 24 hr in both cultures on plastic and basement membrane. Likewise, Prlh antigen was expressed within 24 hr as a homogeneous cytoplasmic fluorescence leaving the basement membrane zone unstained. In contrast, pemphigoid antigen was expressed as a linear fluorescent band at the basement membrane zone between days 3 and 4 of culture. Systematic cell counts of bullous pemphigoid antigen positive cells from trypsin disrupted primary cultures made on plastic over time showed a nadir (8%) of positive cells in early cultures which the percentage rapidly rose to a peak of 58% between days 14 and 21 culture. In subcultures repeatedly disrupted at short intervals, the percentage of bullous pemphigoid positive cells remained low when compared to those interrupted and passaged over longer intervals. The percentage of bullous pemphigoid antigen bearing cells in culture over time is similar, but not identical, to the percentage of basal cells and is related to the age and known growth kinetics of the culture system. Bullous pemphigoid, pemphigus and Prlh antigens are synthesized by the epidermal cell whether cultured on basement membrane or plastic.