Table_4_C-type lectin receptor expression is a hallmark of neutrophils infiltrating the skin in epidermolysis bullosa acquisita.xlsx

IntroductionInflammatory epidermolysis bullosa acquisita (EBA) is characterized by a neutrophilic response to anti-type VII collagen (COL7) antibodies resulting in the development of skin inflammation and blistering. The antibody transfer model of EBA closely mirrors this EBA phenotype.MethodsTo better understand the changes induced in neutrophils upon recruitment from peripheral blood into lesional skin in EBA, we performed single-cell RNA-sequencing of whole blood and skin dissociate to capture minimally perturbed neutrophils and characterize their transcriptome.ResultsThrough this approach, we identified clear distinctions between circulating activated neutrophils and intradermal neutrophils. Most strikingly, the gene expression of multiple C-type lectin receptors, which have previously been reported to orchestrate host defense against fungi and select bacteria, were markedly dysregulated. After confirming the upregulation of Clec4n, Clec4d, and Clec4e in experimental EBA as well as in lesional skin from patients with inflammatory EBA, we performed functional studies in globally deficient Clec4e−/− and Clec4d−/− mice as well as in neutrophil-specific Clec4n−/− mice. Deficiency in these genes did not reduce disease in the EBA model.DiscussionCollectively, our results suggest that while the upregulation of Clec4n, Clec4d, and Clec4e is a hallmark of activated dermal neutrophil populations, their individual contribution to the pathogenesis of EBA is dispensable.

引言：获得性炎症性表皮松解症（EBA）以对抗VII型胶原（COL7）抗体的中性粒细胞反应为特征，导致皮肤炎症和疱状病变的产生。EBA的抗体转移模型与这一EBA表型密切相关。方法：为了更好地理解EBA中从外周血招募至病变皮肤中的中性粒细胞所诱发的变化，我们对全血和皮肤分离物进行了单细胞RNA测序，以捕捉到最小程度受扰动的中性粒细胞并对其转录组进行表征。结果：通过这种方法，我们发现了循环活化中性粒细胞和真皮内中性粒细胞之间的明显差异。最引人注目的是，先前报道的参与宿主对真菌和某些细菌的防御的多聚C型凝集素受体基因表达发生了显著失调。在确认了实验性EBA以及炎症性EBA患者的病变皮肤中Clec4n、Clec4d和Clec4e的上调后，我们在全球性Clec4e−/−和Clec4d−/−小鼠以及中性粒细胞特异性Clec4n−/−小鼠中进行了功能研究。这些基因的缺失并未减少EBA模型中的疾病。讨论：总体而言，我们的结果表明，尽管Clec4n、Clec4d和Clec4e的上调是活化真皮中性粒细胞群体的标志，但它们对EBA发病机制的个体贡献是可替代的。