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Single-cell transcriptomics combined with interstitial fluid proteomics defines cell type-specific immune regulation in atopic dermatitis.
Journal of Allergy and Clinical Immunology ( IF 14.2 ) Pub Date : 2020-04-25 , DOI: 10.1016/j.jaci.2020.03.041
Thomas B Rojahn 1 , Vera Vorstandlechner 2 , Thomas Krausgruber 3 , Wolfgang M Bauer 1 , Natalia Alkon 1 , Christine Bangert 1 , Felix M Thaler 1 , Farzaneh Sadeghyar 1 , Nikolaus Fortelny 3 , Victoria Gernedl 3 , Katharina Rindler 1 , Adelheid Elbe-Bürger 1 , Christoph Bock 4 , Michael Mildner 1 , Patrick M Brunner 1
Affiliation  

Background

Atopic dermatitis (AD) is the most common chronic inflammatory skin disease, but its complex pathogenesis is only insufficiently understood, resulting in still limited treatment options.

Objective

We sought to characterize AD on both transcriptomic and proteomic levels in humans.

Methods

We used skin suction blistering, a painless and nonscarring procedure that can simultaneously sample skin cells and interstitial fluid. We then compared results with conventional biopsies.

Results

Suction blistering captured epidermal and most immune cells equally well as biopsies, except for mast cells and nonmigratory CD163+ macrophages that were only present in biopsy isolates. Using single-cell RNA sequencing, we found comparable transcriptional profiles of key inflammatory pathways between blister and biopsy AD, but suction blistering was superior in cell-specific resolution for high-abundance transcripts (KRT1/KRT10, KRT16/KRT6A, S100A8/S100A9), which showed some background signals in biopsy isolates. Compared with healthy controls, we found characteristic upregulation of AD-typical cytokines such as IL13 and IL22 in Th2 and Th22 cells, respectively, but we also discovered these mediators in proliferating T cells and natural killer T cells, that also expressed the antimicrobial cytokine IL26. Overall, not T cells, but myeloid cells were most strongly enriched in AD, and we found dendritic cell (CLEC7A, amphiregulin/AREG, EREG) and macrophage products (CCL13) among the top upregulated proteins in AD blister fluid proteomic analyses.

Conclusion

These data show that by using cutting-edge technology, suction blistering offers several advantages over conventional biopsies, including better transcriptomic resolution of skin cells, combined with proteomic information from interstitial fluid, unraveling novel inflammatory players that shape the cellular and proteomic microenvironment of AD.



中文翻译:

单细胞转录组学与间质液蛋白质组学相结合定义了特应性皮炎中细胞类型特异性的免疫调节。

背景

特应性皮炎(AD)是最常见的慢性炎症性皮肤病,但对其复杂的发病机理了解甚少,导致治疗选择仍然有限。

目的

我们试图在人类的转录组和蛋白质组学水平上表征AD。

方法

我们使用了皮肤吸水起泡,这是一种无痛且无疤痕的方法,可以同时采样皮肤细胞和间质液。然后,我们将结果与常规活检进行了比较。

结果

抽吸起泡可以捕获表皮细胞和大多数免疫细胞,与活检组织一样好,除了肥大细胞和非迁移性CD163 +巨噬细胞(仅存在于活检分离物中)。使用单细胞RNA测序,我们发现了水泡和活检AD之间关键炎症途径的可比转录谱,但对于高丰度转录本(KRT1 / KRT10,KRT16 / KRT6A,S100A8 / S100A9),吸水泡在细胞特异性分辨率上要好,在活检分离物中显示了一些背景信号。与健康对照相比,我们发现AD常见细胞因子(例如IL 13IL22)在T h 2和T h中具有特征性上调分别为22个细胞,但我们还在增殖T细胞和自然杀伤性T细胞中发现了这些介体,它们也表达了抗菌素细胞因子IL26。总体而言,不是T细胞,而是髓样细胞在AD中的富集程度最高,我们在AD泡罩液蛋白质组学分析中发现树突状细胞(CLEC7A,双调蛋白/ AREG,EREG)和巨噬细胞产物(CCL13)位于最上调的蛋白之中。

结论

这些数据表明,与传统的活组织检查相比,吸塑起泡技术提供了一些优势,包括更好的皮肤细胞转录组学解析,结合组织液中的蛋白质组信息,揭示了塑造AD细胞和蛋白质组微环境的新型炎症因子。

更新日期:2020-04-25
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