Lee CM, et al. Ann Dermatol. 2019 Jun;31(3):349-352. https://doi.org/10.5021/ad.2019.31.3.349

Immunologists have activated T cells in vitro using various stimulation methods, including phorbol myristate acetate (PMA)/ionomycin and αCD3/αCD28 agonistic antibodies. PMA stimulates protein kinase C, activating nuclear factor-κB, and ionomycin increases intracellular calcium levels, resulting in activation of nuclear factor of activated T cell. In contrast, αCD3/αCD28 agonistic antibodies activate T cells through ZAP-70, which phosphorylates linker for activation of T cell and SH2-domain-containing leukocyte protein of 76 kD. However, despite the use of these two different in vitro T cell activation methods for decades, the differential effects of chemical-based and antibody-based activation of primary human T cells have not yet been comprehensively described. Using single-cell RNA sequencing (scRNA-seq) technologies to analyze gene expression unbiasedly at the single-cell level, we compared the transcriptomic profiles of the non-physiological and physiological activation methods on human peripheral blood mononuclear cell–derived T cells from four independent donors. Remarkable transcriptomic differences in the expression of cytokines and their respective receptors were identified. We also identified activated CD4 T cell subsets (CD55+) enriched specifically by PMA/ionomycin activation. We believe this activated human T cell transcriptome atlas derived from two different activation methods will enhance our understanding, highlight the optimal use of these two in vitro T cell activation assays, and be applied as a reference standard when analyzing activated specific disease-originated T cells through scRNA-seq.

We investigated the role of the epithelium of nodules in Hurley stage I hidradenitis suppurativa (HS) by using spatial transcriptomics to analyse the profiles of epithelial cells and dermal-infiltrating immune cells. Compared with epidermal cysts, genes related to bacterial response, inflammatory mediators and neutrophil degranulation pathways were upregulated in the epithelial cells of early-stage HS nodules. Our analysis of dermal-infiltrating immune cells surrounding the epithelium of nodules revealed significantly elevated levels of B-cell-related genes. Similarly to the sinus tract formation observed in moderate-to-severe HS, we propose that the production of inflammatory mediators in early-stage HS may involve the activation of keratinocytes and their interaction with dermal-infiltrating immune cells.