Department of Dermatology, the Second Affiliated Hospital of Kunming Medical University, Kunming 65010, China.
Department of Dermatology, the Second Affiliated Hospital of Kunming Medical University, Kunming 65010, China.
Mol Immunol. 2022 Oct;150:114-125. doi: 10.1016/j.molimm.2022.08.007. Epub 2022 Aug 26.
Pemphigus vulgaris (PV) is a chronic inflammatory autoimmune blistering disease. Aberrant SOCS3/STAT pathway activation is associated with many autoimmune diseases. This study explored the relationship between activation of the SOCS3/STAT pathway and abnormally increased proportions of Th1 and Th17 cells in the peripheral blood of PV patients as well as the effect of CD4 T cells with abnormal SOCS3/STAT pathway activation on acantholysis.
In PV patients, the proportions of Th1 and Th17 cells in peripheral blood, the levels of IFN-γ and IL-17 in serum and the mRNA levels of SOCS3 and STAT1/3 in CD4 T cells were detected. Then, SOCS3-knockdown primary CD4 T cells were prepared, and cocultured with HaCaT cells. Finally, after SOCS3 knockdown and coculture, CD4 T cells were collected, and the proportions of Th1 and Th17 cells, the protein levels of STAT1/3 and p-STAT1/3, and the levels of IFN-γ and IL-17 were measured. After 2 days of coculture, HaCaT cells were collected, inflammatory factors mRNA expression and acantholysis were assessed.
In PV patients, the proportions of Th1 (P = 0.016) and Th17 (P = 0.045) cells and the levels of IFN-γ (P = 0.010) were significantly increased. SOCS3 mRNA in CD4 T cells was significantly decreased (P = 0.008), whereas STAT1 (P = 0.043) and STAT3 (P = 0.004) mRNA were significantly increased. After SOCS3 knockdown, the proportions of Th1 (P < 0.001) and Th17 (P = 0.006) cells, the levels of IFN-γ (P < 0.001) and IL-17 (P = 0.001), and the protein levels of p-STAT1 (P = 0.001) and p-STAT3 (P = 0.003) were significantly increased in the CD4 T-shSOCS3-1 group. In the coculture system, the proportions of Th1 (P < 0.001) and Th17 (P < 0.001) cells, the levels of IFN-γ (P < 0.001) and IL-17 (P < 0.001), and the number of cell fragments (P < 0.001) were significantly increased in the CD4 T-shSOCS3-1+HaCaT-PV-IgG group, whereas the protein level of desmoglein3 (Dsg3) was significantly decreased. In addition, PV-IgG significantly increased IFN-γ and IL-6 mRNA in HaCaT cells.
Low SOCS3 expression in CD4 T cells from PV patients leads to overactivation of STAT, which causes CD4 T cells to overdifferentiate into Th1 and Th17 cells. Additionally, PV-IgG-induced local inflammation in skin lesions, which is mediated by IFN-γ and IL-6, can aggravate this phenomenon. Furthermore, low SOCS3 expression in CD4 T cells further exacerbates PV-IgG-induced acantholysis. Therefore, upregulating the expression of SOCS3 in CD4 T cells of PV patients and maintaining the balance of the IFN-γ/STAT1/SOCS3 and IL-6/STAT3/SOCS3 pathways can alleviate acantholysis in patients with PV.
寻常型天疱疮（PV）是一种慢性炎症性自身免疫性水疱病。异常的 SOCS3/STAT 通路激活与许多自身免疫性疾病有关。本研究探讨了 SOCS3/STAT 通路的激活与 PV 患者外周血中 Th1 和 Th17 细胞比例异常增加之间的关系，以及异常激活 SOCS3/STAT 通路的 CD4 T 细胞对棘层松解的影响。
检测 PV 患者外周血中 Th1 和 Th17 细胞的比例、血清中 IFN-γ和 IL-17 的水平、CD4 T 细胞中 SOCS3 和 STAT1/3 的 mRNA 水平。然后，制备 SOCS3 敲低的原代 CD4 T 细胞，并与 HaCaT 细胞共培养。最后，在 SOCS3 敲低和共培养后，收集 CD4 T 细胞，检测 Th1 和 Th17 细胞的比例、STAT1/3 和 p-STAT1/3 的蛋白水平、IFN-γ和 IL-17 的水平。共培养 2 天后，收集 HaCaT 细胞，评估炎症因子 mRNA 表达和棘层松解情况。
PV 患者 Th1（P=0.016）和 Th17（P=0.045）细胞比例及 IFN-γ水平显著升高（P=0.010）。CD4 T 细胞中 SOCS3 mRNA 显著降低（P=0.008），而 STAT1（P=0.043）和 STAT3（P=0.004）mRNA 显著升高。SOCS3 敲低后，CD4 T-shSOCS3-1 组 Th1（P<0.001）和 Th17（P=0.006）细胞比例、IFN-γ（P<0.001）和 IL-17（P=0.001）水平、p-STAT1（P=0.001）和 p-STAT3（P=0.003）蛋白水平显著升高。在共培养系统中，CD4 T-shSOCS3-1+HaCaT-PV-IgG 组 Th1（P<0.001）和 Th17（P<0.001）细胞比例、IFN-γ（P<0.001）和 IL-17（P<0.001）水平、细胞碎片数量（P<0.001）均显著增加，而桥粒芯糖蛋白 3（Dsg3）蛋白水平显著降低。此外，PV-IgG 可显著增加 HaCaT 细胞中 IFN-γ和 IL-6 mRNA 的表达。
PV 患者 CD4 T 细胞中 SOCS3 表达降低导致 STAT 过度激活，使 CD4 T 细胞过度分化为 Th1 和 Th17 细胞。此外，PV-IgG 诱导皮肤病变局部炎症，这是由 IFN-γ和 IL-6介导的，可加重这种现象。此外，CD4 T 细胞中 SOCS3 表达降低进一步加重了 PV-IgG 诱导的棘层松解。因此，上调 PV 患者 CD4 T 细胞中 SOCS3 的表达，维持 IFN-γ/STAT1/SOCS3 和 IL-6/STAT3/SOCS3 通路的平衡，可以减轻 PV 患者的棘层松解。
