Abstract
ABSTRACTThe 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.
Highlights
The immune system constantly surveys the microenvironment and discriminates between harmless and potentially harmful components – a multifaceted task at barrier surfaces, such as the skin, that are constantly exposed to exogenous stimuli
T cell yields were comparatively assessed from late second-trimester fetal and adult skin biopsies using established as well as novel isolation techniques and subsequent flow cytometry and confocal microscopy analyses (Fig. 1A,B)
In this study, we used a combination of advanced approaches to further explore the complexity and heterogeneity of T cell subsets in fetal human skin
Summary
The immune system constantly surveys the microenvironment and discriminates between harmless and potentially harmful components – a multifaceted task at barrier surfaces, such as the skin, that are constantly exposed to exogenous stimuli. There, a complex network of cellular and molecular pathways is employed, T.A., 0000-0001-6275-4607; J.S., 0000-0003-1605-2393; P.K., 0000-0002-01146904; N.F., 0000-0003-4025-9968; P.A.V.-G., 0000-0001-7101-9313; C.W., 00000002-7635-3721; C.F., 0000-0001-6451-7349; W.E., 0000-0001-9669-7989; A.S., 0000-0001-7368-3226; T.K., 0000-0002-1374-0329; A.E.-B., 0000-0003-24610367 Handling Editor: Paul Martin Received 5 May 2021; Accepted 7 September 2021 which allows the immune system to respond quickly and efficiently to harmful stimuli, while largely ignoring innocuous substances. Regulation of both innate and adaptive skin immunity is essential in preserving host integrity, thereby preventing inappropriate immune activation and pathology. Multiple T cell subsets are involved in the defense against pathogens and tumors, and play a role in tissue homeostasis (e.g. hair follicle cycling, wound repair), but they can cause inflammation and autoimmune diseases (Cruz et al, 2018; Ho and Kupper, 2019)
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