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. 2025 Jun 20;193(1):115-124.
doi: 10.1093/bjd/ljaf041.

Comprehensive single-cell chromatin and transcriptomic profiling of peripheral immune cells in nonsegmental vitiligo

Jesus Gay-Mimbrera  1 , Daniel Lozano-Ojalvo  2 , Pedro J Gómez-Arias  1   3 , Irene Rivera-Ruiz  3 , Macarena Aguilar-Luque  1 , Carmen Mochón-Jiménez  1   3 , Eloísa Andújar Pulido  4 , Mónica Pérez-Alegre  4 , Emma Guttman-Yassky  2 , Juan Ruano  1   3
Affiliations

Comprehensive single-cell chromatin and transcriptomic profiling of peripheral immune cells in nonsegmental vitiligo

Jesus Gay-Mimbrera et al. Br J Dermatol. .

Abstract

Background: Nonsegmental vitiligo (NSV) is an autoimmune condition characterized by melanocyte loss. While skin-specific mechanisms have been well studied, systemic immune dysregulation contributing to NSV pathogenesis remains unclear.

Objectives: To use a multi-omic single-cell approach to investigate circulating immune cells in NSV, integrating transcriptional and chromatin accessibility data.

Methods: An integrative single-cell RNA sequencing (scRNAseq)/single-cell assay for transposase-accessible chromatin sequencing (scATACseq) analysis was conducted on peripheral blood mononuclear cells (PBMCs) from people with NSV (n = 11) and healthy control participants (n = 5), identifying transcriptional markers, cell-cell interactions, chromatin accessibility and transcription factor (TF) dynamics. Key findings were validated in an expanded cohort (patients with NSV, n = 16; healthy controls, n = 9) using spectral flow cytometry, with additional stratification by sex, age, disease activity, severity and duration.

Results: Analysis of 59 192 PBMCs identified 8204 gene expression markers and 13 925 ATAC peaks across 25 immune cell subtypes. A broadly activated immune response was observed, characterized by cytotoxicity, antigen presentation, cell exhaustion and stress, predominantly in monocytes, natural killer cells, CD8+ T cells and dendritic cells. Multi-omic integration revealed T helper (Th)1/Th17 polarization and dysfunctional regulatory T cell [Treg/memory Treg (mTreg)] responses. Chromatin accessibility highlighted enriched TF binding sites for forkhead box O3 (FOXO3), Sp1, activator protein 1 (AP-1), signal transducer and activator of transcription (STAT)1/STAT3, interferon regulatory factor (IRF)1 and IRF4, regulating pathways linked to cytotoxicity, antigen processing, nuclear factor-κB, Toll-like receptor and Janus kinase/STAT signalling. Flow cytometry validated these findings, showing that disease activity and shorter duration were associated with heightened immune dysregulation. Robust T-cell receptor activation drove Th1/Th17 polarization and elevated interferon-γ and tumour necrosis factor-α production in CD4+ and CD8+ T cells. Cutaneous lymphocyte-associated antigen (CLA)+ skin-homing Th1/Th17-polarized CD4+ T cells, CD8+ T cells and mTregs exhibited persistent activation, marked by basal programmed cell death protein 1 (PD1)+ expression. OX40/OX40L-mediated interactions between monocytes and effector T cells amplified inflammation. Regulatory dysfunction, including reduced interleukin (IL)-4 and IL-13 production by mTregs, was prominent in moderate-to-severe and active disease.

Conclusions: This is the first multi-omic single-cell study of PBMCs from people with NSV, revealing systemic immune dysregulation driven by cytotoxicity, antigen presentation, exhaustion and regulatory failure. Disease severity, activity and evolution influence these pathways, highlighting the OX40/OX40L axis as a potential therapeutic target to mitigate immune dysregulation and relapse risk.

Plain language summary

Vitiligo is a common condition that causes patches of white skin. This happens because the immune system mistakenly attacks the cells responsible for skin colour. These cells are called ‘melanocytes’. Non-segmental vitiligo (‘NSV’ for short) is the most common form and can spread across the body over time. In this study, researchers from Spain and the USA looked at how the immune system contributes to NSV. We examined immune cells circulating in the blood of people with NSV. Using advanced techniques, we looked at gene activity and DNA structure in these cells. This approach helped us identify which genes and pathways might cause immune cells to attack skin cells. The results showed that specific immune cells were very active. They also showed signs of inflammation and stress, leading to an aggressive immune response. We also found that some immune cells were not functioning in the right way. A key finding was that a specific pathway involving proteins called ‘OX40’ and ‘OX40L’ could drive this imbalance in the immune system. Future treatments could use this pathway to stop the immune system from attacking the skin’s pigment cells. It could also reduce disease progression. This research doesn’t change treatments right now, but it could lead to new and better treatments for people with vitiligo in the future.

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Conflict of interest statement

Conflicts of interest: The authors declare no conflicts of interest.

Comment in

  • Focusing on antigen-specific T cells.
    Inozume T, Fukushima S. Inozume T, et al. Br J Dermatol. 2025 Jun 20;193(1):6-7. doi: 10.1093/bjd/ljaf050. Br J Dermatol. 2025. PMID: 39918099 No abstract available.