Department of Dermatology, Medical University of Vienna, Vienna 1090, Austria.
Department of Biosciences, University of Salzburg, Salzburg 5020, Austria.
Development. 2022 Apr 15;149(8). doi: 10.1242/dev.199781. Epub 2021 Oct 26.
The adult human skin contains a vast number of T cells that are essential for skin homeostasis and pathogen defense. T cells are first observed in the skin at the early stages of gestation; however, our understanding of their contribution to early immunity has been limited by their low abundance and lack of comprehensive methodologies for their assessment. Here, we describe a new workflow for isolating and expanding significant amounts of T cells from fetal human skin. Using multiparametric flow cytometry and in situ immunofluorescence, we found a large population with a naive phenotype and small populations with a memory and regulatory phenotype. Their molecular state was characterized using single-cell transcriptomics and TCR repertoire profiling. Importantly, culture of total fetal skin biopsies facilitated T cell expansion without a substantial impact on their phenotype, a major prerequisite for subsequent functional assays. Collectively, our experimental approaches and data advance the understanding of fetal skin immunity and potential use in future therapeutic interventions.
成人皮肤中含有大量的 T 细胞，这些细胞对于皮肤的稳态和病原体防御至关重要。T 细胞最早在妊娠早期的皮肤中被观察到；然而，由于它们的丰度低，以及缺乏全面的评估方法，我们对它们在早期免疫中的贡献的理解一直受到限制。在这里，我们描述了一种从胎儿皮肤中分离和扩增大量 T 细胞的新工作流程。使用多参数流式细胞术和原位免疫荧光技术，我们发现了一个具有幼稚表型的大群体，以及具有记忆和调节表型的小群体。使用单细胞转录组学和 TCR 库分析对其分子状态进行了表征。重要的是，总胎儿皮肤活检的培养促进了 T 细胞的扩增，而对其表型没有显著影响，这是后续功能测定的主要前提。总的来说，我们的实验方法和数据推进了对胎儿皮肤免疫的理解，并为未来的治疗干预提供了潜在的应用。
