Imaging Mass Cytometry (IMC) allows for the visualisation of up to 40 biomarkers on a single tissue section via the use of metal-tagged antibodies, permitting advanced multiplex imaging for detailed in situ analyses. To test whether this technique could be established to enhance the study of inflammatory hair diseases, we tested and optimised a panel of tissue architecture, immune infiltration, and immune activation markers on non-lesional and lesional alopecia areata (AA) frozen tissue sections. Analysis of data obtained via the Hyperion Imaging System revealed successful multiplex staining on both non-lesional and lesional AA tissue. Specifically, the hair follicle (HF) bulb was stained for Ki-67, epithelial E-Cadherin and Pan-Keratin, and mesenchymal Vimentin, Collagen Type 1, and Alpha-smooth muscle actin. Consistent with previous reports, in lesional AA HFs the inflammatory infiltrate around the bulb was successfully labelled with markers of helper and cytotoxic T Cells (CD3, CD4 and CD8), memory and regulatory T cells (CD45RO, FOXP3) and macrophages (CD68). Granzyme B was also labelled in the lesional AA bulb. This work highlights the potential of IMC to readily multiplex several markers highly relevant to hair pathology, leaving ample scope to further optimise and complement this panel with additional markers of interest (e.g., pertaining to immune privilege or immunometabolism). In conclusion, this technique provides a powerful new means to perform enhanced phenotyping of the HF and to analyse novel immune cell and tissue interactions not only in AA, but in other hair pathologies such as scarring alopecia.

ESDR368 - Methods in Inflammatory Hair Disease Research: Enhanced Multiplex Imaging on Alopecia Areata Tissue Sections

The scientific meeting of the European Society for Dermatological Research (ESDR) is held yearly, usually in September. With an international attendance of more than 1000 skin scientists from around the globe, the four-day event includes short oral presentations from peer-reviewed abstracts, keynote lectures from international experts, sponsored symposia, satellite programs, educational events as well as an important focus on posters, networking and building collaborations amongst scientists. For more information about this year’s meeting visit our website.