背景:表皮松解性鱼鳞病(EI)是非综合征性遗传性鱼鳞病的主要形式,以红皮病为特征,明显的角化过度和鳞屑,bulla和出生时的侵蚀,与KRT1/KRT10突变相关。EI中的细胞因子和趋化因子谱知之甚少,和具体的治疗方案尚未确定。
目的:探索EI患者的新型生物标志物和治疗靶点。
方法:我们分析了10例遗传性鱼鳞病患者血清和皮肤样本中的细胞因子水平,包括7名EI患者.建立野生型和突变型KRT1构建体,转染HaCaT细胞,一种永生化的角化细胞系,用于体外免疫印迹和免疫细胞化学分析。
结果:多重细胞因子/趋化因子分析显示10种细胞因子/趋化因子[白细胞介素(IL)-1β,IL-4,IL-17A,IL-16,IL-18,IL-1受体-α,巨噬细胞集落刺激因子,EI患者的干扰素-α2,碱性成纤维细胞生长因子和单核细胞趋化蛋白-3]显着增加。此外,EI患者的IL-18水平[n=7;2714.1(1438.0)pgmL-1]明显高于健康对照组[n=11;218.4(28.4)pgmL-1,P<0.01]。免疫组织化学分析显示,EI患者皮肤样本中IL-18表达升高。血清IL-18水平与鱼鳞病的严重程度相关,由鱼鳞病评分系统测量。免疫印迹分析显示,在表达突变KRT1的HaCaT细胞的上清液中成熟IL-18水平增加。此外,这些细胞在细胞质中显示NLRP3聚集,ASC聚集在突变的角蛋白聚集周围.这些发现表明,突变角蛋白可能会促进EI患者角质形成细胞中NLRP3炎性体的激活及其下游caspase-1介导的IL-18释放。
结论:我们的结果表明,血清IL-18是从EI患者皮肤释放的严重程度标志物。IL-18的阻断可能是EI患者的一种有用的新型治疗选择。
BACKGROUND: Epidermolytic ichthyosis (EI) is a major form of nonsyndromic inherited ichthyosis, characterized by erythroderma, marked hyperkeratosis and scale, bulla and erosion at birth, associated with KRT1/KRT10 mutations. The cytokine and chemokine profiles in EI are poorly understood, and specific treatment options have not been established.
OBJECTIVE: To explore novel biomarkers and therapeutic targets in patients with EI.
METHODS: We analysed cytokine levels in serum and skin samples from 10 patients with inherited ichthyosis, including seven patients with EI. Wild-type and mutant KRT1 constructs were established and transfected into HaCaT cells, an immortalized keratinocyte cell line, for in vitro immunoblotting and immunocytochemistry analyses.
RESULTS: Multiplex cytokine/chemokine analysis revealed that 10 cytokines/chemokines [interleukin (IL)-1β, IL-4, IL-17A, IL-16, IL-18, IL-1 receptor-α, macrophage colony-stimulating factor, interferon-α2, basic fibroblast growth factor and monocyte chemotactic protein-3] were significantly increased in patients with EI. Furthermore, IL-18 levels were significantly higher in patients with EI [n = 7; 2714.1 (1438.0) pg mL-1] than in healthy controls [n = 11; 218.4 (28.4) pg mL-1, P < 0.01]. Immunohistochemical analyses showed that IL-18 expression was elevated in skin samples from patients with EI. Serum IL-18 levels correlated with the severity of ichthyosis, as measured by the Ichthyosis Scoring System. Immunoblotting analysis revealed that mature IL-18 levels were increased in the supernatant of mutant KRT1 expressing HaCaT cells. Additionally, these cells showed NLRP3 aggregation in the cytoplasm and ASC clustered around mutant keratin aggregations. These findings suggest that mutant keratin might promote the activation of the NLRP3 inflammasome and its downstream caspase-1-mediated IL-18 release in keratinocytes from patients with EI.
CONCLUSIONS: Our results suggest that serum IL-18 is a severity marker released from the skin of patients with EI. Blockade of IL-18 may be a useful novel therapeutic option for patients with EI.