Impact of innate immunity on pemphigus vulgaris disease manifestation

如银屑病所示，先天免疫在T细胞介导的疾病中起着重要作用。因此，我们旨在探讨先天免疫系统在抗体介导的自身免疫性疾病寻常型天疱疮中的作用。尽管PV依赖于抗Dsg3特异性抗体的发展，但疾病的表现需要额外的因素。其中一个因素可能是先天免疫系统。先前在天疱疮小鼠模型中的发现和在天疱疮患者中的体外研究结果表明，由IL-1诱导的IL-1和IL-6在自身抗体诱导的表皮内粘连丧失中起关键作用。在证明了先天免疫的细胞因子可由PV抗体、机械应力和紫外线诱导后，我们使用3D皮肤模型用PV IgG处理角质形成细胞和成纤维细胞，并将辅因子局部地添加到表皮或3D皮肤等效物的液体中。此外，我们将在系统中加入巨噬细胞和中性粒细胞，以模拟细胞的先天渗透。使用转基因角质形成细胞或药物抑制，将评估棘层松解对NFκB、IL-1、NLR或Caspase存在的功能依赖性。这一方法可以阐明水泡形成的主要先天途径，并消除基底部以上棘层松解的机制。此外，桥粒蛋白-1的缺失或DSG1的突变与SAM综合征有关，DSG1是在叶天疱疮中产生抗体的结构。这种罕见的疾病与严重的消瘦、经皮水分丢失和IL-1细胞因子的产生有关。除了天然免疫对水泡形成的影响外，另一个目的是破译DSG1对角质形成细胞，特别是PV中炎症反应的影响。我们将使用3D皮肤模型来治疗它们，要么用抗DSG1抗体，要么用DSG1缺陷或突变的角质形成细胞。然后，我们的目标是研究一种不同的炎症模式，这可能解释SAM综合征和PF的不同表型。由于DSG1的存在对抗Dsg3诱导的棘层松解有保护作用，因此我们将研究DSG-1缺乏时炎症信号对PV的影响。在这里，我们利用最近建立的粘膜3D-皮肤模型，因为粘膜上皮不携带大量的DSG1。建议的实验将阐明先天免疫和DSG1在棘层松解和PV表现中的作用。由于IL-1和IL-6通路在药理学上都是靶向性的，这可能导致识别新的靶点，可以用于有效的治疗。

As shown for psoriasis, innate immunity can serve an important role in T cell mediated diseases. Therefore, we aim to investigate the impact of the innate immune system on the antibody mediated autoimmune disease pemphigus vulgaris. Even though PV relies on the development of anti-Dsg3-specific antibodies, disease manifestation requires additional factors. One of these factors might be the innate immune system. Previous findings in pemphigus mouse models and ex vivo findings in pemphigus patients suggest that IL-1 and IL-6 which is induced by IL-1 are critical for auto-ab-induced intraepidermal loss of adhesion. After having shown that cytokines of innate immunity are induced by PV antibodies, by mechanical stress, and by UV, we use 3D-skin models to treat keratinocytes and fibroblasts with PV IgG and add cofactors either topically on the epidermis or to the liquid phase of the 3D-skin equivalents. In addition, we will include macrophages and neutrophils in the system to mimic cellular innate infiltration. Using genetically modified keratinocytes or pharmacological inhibition, the functional dependency of acantholysis on the presence of NFκB, IL-1, NLRs or caspases will be evaluated. This approach can elucidate the responsible innate pathway for blister formation and dismantle the mechanism of suprabasal acantholysis. Furthermore, a loss of desmoglein-1 or mutations in Dsg1, the structure against which antibodies are generated in pemphigus foliaceus, are associated with SAM-syndrome. This rare disease is associated with severe wasting, transepidermal water-loss and the production of IL-1 cytokines. A further aim besides the impact of innate immunity on blister formation ist to decipher the impact of Dsg1 on the inflammatory response in keratinocytes and in particular in PV. We will use 3D-skin models to either treat them with anti-Dsg1 or with Dsg1-deficient or mutated keratinocytes. Then we aim to investigate a different inflammatory pattern which might explain the different phenotypes in SAM-syndrome and PF. As the presence of Dsg1 protects from anti-Dsg3 induced acantholysis, we will then investigte the impact of the inflammatory signature in Dsg-1 deficiency on PV. Here, we utilize a recently established mucous membrane 3D-skin model as the mucosal epithelia do not carry significant amounts of Dsg1. The proposed experiments will clarify the role of the innate immune and of Dsg1 in acantholysis and PV manifestation. As both the IL-1 and IL-6 pathways are pharmacologically targetable, this might lead to the identification of new targets that could be used as effective therapy.