Department of Immunology, School of Biological Sciences, UACB, Universidad Autónoma de Zacatecas, Av. de la Revolución Mexicana s/n, Colonia Tierra y Libertad, Guadalupe CP 98615, Zacatecas, Mexico.
School of Chemistry Sciences, Universidad Autónoma de Zacatecas, Campus Universitario Siglo XXI, Carretera Zacatecas-Guadalajara, Ejido "La Escondida", Zacatecas CP 98160, Zacatecas, Mexico.
Int J Mol Sci. 2024 Aug 14;25(16):8864. doi: 10.3390/ijms25168864.
Pemphigus is an autoimmune disease that affects the skin and mucous membranes, induced by the deposition of pemphigus IgG, which mainly targets desmogleins 1 and 3 (Dsg1 and 3). This autoantibody causes steric interference between Dsg1 and 3 and the loss of cell adhesion, producing acantholysis. This molecule and its cellular effects are clinically reflected as intraepidermal blistering. Pemphigus vulgaris-IgG (PV-IgG) binding involves p38MAPK-signaling-dependent caspase-3 activation. The present work assessed the in vitro effect of PV-IgG on the adherence of cells dependent on caspase-3. PV-IgG induced cell detachment and apoptotic changes, as demonstrated by annexin fluorescent assays. The effect of caspase-3 induced by PV-IgG was suppressed in cells pre-treated with caspase-3-shRNA, and normal IgG (N-IgG) as a control had no relevant effects on the aforementioned parameters. The results demonstrated that shRNA reduces caspase-3 expression, as measured via qRT-PCR and via Western blot and immunofluorescence, and increases cell adhesion. In conclusion, shRNA prevented in vitro cell detachment and the late effects of apoptosis induced by PV-IgG on cells, furthering our understanding of the molecular role of caspase-3 cell adhesion dependence in pemphigus disease.
天疱疮是一种自身免疫性疾病，影响皮肤和黏膜，由天疱疮 IgG 的沉积引起，主要针对桥粒芯糖蛋白 1 和 3（Dsg1 和 3）。这种自身抗体导致 Dsg1 和 3 之间的空间干扰和细胞黏附丧失，产生棘层松解。这种分子及其细胞作用在临床上表现为表皮内水疱形成。寻常型天疱疮-IgG（PV-IgG）结合涉及 p38MAPK 信号依赖性半胱天冬酶-3 激活。本工作评估了 PV-IgG 对依赖半胱天冬酶-3 的细胞黏附的体外影响。PV-IgG 诱导细胞脱落和凋亡变化，如 Annexin 荧光测定所示。用 caspase-3-shRNA 预处理细胞可抑制 PV-IgG 诱导的 caspase-3 作用，而作为对照的正常 IgG（N-IgG）对上述参数无相关影响。结果表明，shRNA 降低了 caspase-3 的表达，如通过 qRT-PCR 和 Western blot 及免疫荧光测定所示，并增加了细胞黏附。综上所述，shRNA 可预防体外细胞脱落和 PV-IgG 诱导的细胞凋亡的晚期效应，进一步了解了半胱天冬酶-3 对天疱疮疾病中细胞黏附的分子作用。
