The molecular and phenotypic makeup of fetal human skin T lymphocytes

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.

Automated summary

This study describes a new workflow for isolating and expanding significant amounts of T cells from fetal human skin. The characteristics of T cells in fetal skin were determined using multiparametric flow cytometry and immunofluorescence techniques. The molecular state of these cells was characterized using single-cell transcriptomics and TCR repertoire profiling.