Woodley D, Saurat J H, Pruniéras M, Régnier M
J Invest Dermatol. 1982 Jul;79(1):23-9. doi: 10.1111/1523-1747.ep12510438.
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 was 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 after which the percentage rapidly rose to a peak of 58% between days 14 and 21 of 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 cultures system. Bullous pemphigoid, pemphigus and Prlh antigens are synthesized by the epidermal cell whether cultured on basement membrane or plastic.
将分离出的成人人类角质形成细胞培养在塑料盖玻片上或含有完整层粘连蛋白、IV型和V型胶原蛋白以及硫酸乙酰肝素蛋白聚糖的无活力基底膜表面，通过间接免疫荧光检查大疱性类天疱疮、天疱疮和Prlh抗原的表达。初始细胞悬液中，大疱性类天疱疮和Prlh阳性染色细胞的平均比例分别为23%和30%，而用天疱疮血清染色的细胞通常为阴性（22个系列中有19个）。在塑料和基底膜上的两种培养物中，天疱疮抗原在24小时内均表现为角质形成细胞间的细胞内染色。同样，Prlh抗原在24小时内表现为均匀的细胞质荧光，基底膜区未染色。相比之下，类天疱疮抗原在培养的第3至4天表现为基底膜区的线性荧光带。随着时间的推移，对胰蛋白酶处理的塑料上原代培养物中分离出的大疱性类天疱疮抗原阳性细胞进行系统计数，结果显示早期培养物中阳性细胞的最低点（8%），之后在培养的第14至21天，阳性细胞百分比迅速上升至峰值58%。在短时间间隔反复传代培养中，与较长间隔传代培养相比，大疱性类天疱疮阳性细胞的百分比仍然较低。随着时间的推移，培养物中携带大疱性类天疱疮抗原细胞的百分比与基底细胞的百分比相似但不相同，并且与培养系统的年龄和已知生长动力学有关。无论培养在基底膜还是塑料上，表皮细胞都能合成大疱性类天疱疮、天疱疮和Prlh抗原。
