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目的 利用HaCaT细胞与寻常型天疱疮患者血清IgG (PV-IgG)共培养构建天疱疮细胞模型,并观察棘层松解过程中细胞的变化情况.方法 收集典型寻常型天疱疮患者,从血清中纯化IgG,以1mg/mL的浓度与HaCaT细胞共培养,通过细胞解离实验观察PV-IgG对HaCaT细胞黏附功能的影响;通过免疫荧光观察桥粒蛋白的染色模式;通过免疫印迹观察桥粒分子蛋白质水平的变化;通过定量多聚酶链反应(qPCR)观察桥粒分子mRNA水平的变化;通过免疫共沉淀了解Dsg3与PG的相互作用;通过磷酸化测定观察p38 MAPK、EGFR磷酸化水平.结果 PV-IgG与HaCaT细胞共培养后,可以使HaCaT细胞碎片数目增多,桥粒分子内化降解、稳定性下降,Dsg3与PG相互作用减弱,但对桥粒分子mRNA水平的表达没有影响;细胞内信号分子p38 MAPK与EGFR出现磷酸化,且p38 MAPK的磷酸化发生于EGFR磷酸化之前.结论 利用HaCaT细胞和PV-IgG共培养可以构建天疱疮的细胞模型,模型中的桥粒分子出现了与体内棘层松解过程一致的变化.“,”Objective To establish a cell model of pemphigus using co-cultured HaCaT cells with pemphigus vulgaris (PV)-IgG.Methods Typical patients with pemphigus vulgaris were enrolled.IgG in the serum of the patients was purified.Then HaCaT cells were co-cultured with PV-IgG of concentration of 1mg/mL.The cell dissociation assay was conducted to detect the adhesion function of HaCaT cells and immunofluorescent assay was conducted to observe the staining patterns of desmosome proteins.Western blot was conducted to analyze the protein level of adhesion molecules;qPCR was conducted to detect desmosome mRNA level.The interaction of desmoglein3 with plakoglobin was qualitatively analyzed by co-immunoprecipitation.The phosphorylation levels of p38 MAPK and EGFR were examined.Results After co-culture of HaCaT cells with PV-IgG,the fragments of HaCaT cells increased.The molecules of desmosome protein were internalized and degraded and its stability declined.The interaction between Dsg3 and PG decreased.The expression of desmosome mRNA level was not affected.The phosphorylation of p38 MAPK and EGFR appeared,and the phosphorylation of p38 MAPK occurred earlier than that of EGFR.Conclusion The cell model of pemphigus can be successfully established by co-culture of HaCaT cells with PV-IgG,and desmosome has the consistent changes that the acantholysis process happened in vivo.