不同亚型天疱疮患者血清对HaCaT细胞桥粒芯糖蛋白3表达的影响

Effects of serum from different subtypes of pemphigus on the expression of desmoglein-3 in HaCaT cells

目的:确定3例不同亚型天疱疮患者(寻常型、副肿瘤型和落叶型天疱疮)血清对HaCaT细胞桥粒芯糖蛋白1、3(DSG1、3)及mRNA表达的影响。方法:采集3例患者血清,ELISA测定抗DSG1抗体和抗DSG3抗体;血清培养HaCaT细胞,直接免疫荧光检测细胞表面DSG1、3荧光强度,qPCR测定DSG3 mRNA的表达。结果:寻常型天疱疮患者血清抗DSG1抗体及抗DSG3抗体均阳性,副肿瘤型天疱疮患者血清抗DSG3抗体阳性,落叶型天疱疮患者血清抗DSG1抗体阳性。3例患者血清分别作用后,HaCaT细胞表面DSG3荧光强度均降低;寻常型及副肿瘤型天疱疮患者血清作用后,HaCaT细胞DSG3 mRNA表达增加,落叶型天疱疮患者血清作用后,HaCaT细胞DSG3 mRNA表达降低。结论:抗DSG1抗体血清可以降低DSG3表达;抗DSG3抗体血清可以增加细胞中DSG3的表达,但血清中的抗DSG3抗体存在以及KC细胞对DSG3的内吞等作用会将其表达的蛋白减少。

Objective：To determine the effects of serum from 3 patients with different subtypes of pemphigus （ pemphigus vulgaris, pemphigus foliaceus and paraneoplastic pemphigus） on the expression of desmoglein-3 and mRNA in HaCaT cells. Methods： The levels of anti-DSG1 and anti-DSG3 autoantibodies from patients’ serum were detected by ELISA. HaCaT cells were cultured in serum-DMEM culture. The content of DSG3 protein on HaCaT cells were detected by direct immunofluorescence and DSG3 mRNA were detected by qPCR. Results：ELISA showed positive of anti-DSG1 and anti-DSG3 autoantibodies in the patient with pemphigus vulgaris, positive of anti-DSG3 autoantibodies in the patient with paraneoplastic pemphigus and positive of anti-DSG1 autoantibodies in the patient with pemphigus foliaceus. The fluorescence intensity of DSG3 on the surface of HaCaT cells was decreased in the three patients after separately cultured with the sera of the three patients. The content of DSG3 mRNA in HaCaT cells was increased after cultured with serum of the patients with pemphigus vulgaris and paraneoplastic pemphigus. The content of DSG3 mRNA in HaCaT cells was decreased after cultured with serum of the patient with pemphigus foliaceus. Conclusion： The serum containing anti-DSG1 antibodies decreased the expression of DSG3 mRNA. The serum containing anti-DSG3 anti-bodies increased the expression of DSG3 mRNA but the protein of DSG3 was decreased because of the endocytosis of keratinocytes and existence of anti-DSG3 antibodies in the serum.