Abstract
Generalized pustular psoriasis (GPP) is a severe subtype of psoriasis characterized by epidermal neutrophil infiltration, often presenting as acute, potentially life-threatening flares. However, the characterization of the immune micro-environment in GPP lesions remains largely unknown. Here, we use single-cell RNA profiling to interrogate the transcriptomes of 60,000 single cells from GPP lesional skin (n = 13) and healthy adult skin (n = 4), combined with spatial transcriptomics. We identify a neutrophil subset lacking CASP8 expression but exhibiting elevated levels of inflammatory pathway genes, including RIPK1, NFKB1, IL1B, CXCL1, and CXCL8 in GPP flares, illustrating neutrophil transition from pre-inflammatory to a pro-inflammatory state, and activation of a communication network between IL36G+ keratinocytes and neutrophils in GPP lesions, with TNFSF15 (TL1A) released from neutrophils exaggerating the inflammatory crosstalk. We further demonstrate that fibroblasts and capillary endothelial cells function as central communication hubs in GPP, through dynamic receptor-ligand interactions with several spatially proximate immune cells, including T cells, neutrophils, and macrophages. In this work, we provide an in-depth view of immune cell participation and highlight the role of neutrophil-keratinocyte crosstalk in GPP pathogenesis.
Similar content being viewed by others
Data availability
The scRNA-seq data generated are available in GEO under accession number GSE309097 (GEO Accession viewer). The spatial transcriptomics data generated for GPP samples are available in Zenodo (GPP Xenium). The remaining data are available within the Supplementary Information or Source data file from the corresponding author on request. Source data are provided with this paper.
References
Marrakchi, S. & Puig, L. Pathophysiology of generalized pustular psoriasis. Am. J. Clin. Dermatol. 23, 13–19 (2022).
Rivera-Díaz, R., Daudén, E., Carrascosa, J. M., Cueva, P. & Puig, L. Generalized pustular psoriasis: a review on clinical characteristics, diagnosis, and treatment. Dermatol. Ther. 13, 673–688 (2023).
Onoufriadis, A. et al. Mutations in IL36RN/IL1F5 are associated with the severe episodic inflammatory skin disease known as generalized pustular psoriasis. Am. J. Hum. Genet. 89, 432–437 (2011).
Arakawa, A., Ruzicka, T. & Prinz, J. C. Therapeutic efficacy of interleukin 12/interleukin 23 blockade in generalized pustular psoriasis regardless of IL36RN mutation status. JAMA Dermatol. 152, 825–828 (2016).
Prinz, J. et al. Chimaeric CD4 monoclonal antibody in treatment of generalised pustular psoriasis. Lancet 338, 320–321 (1991).
Arakawa, A. et al. Unopposed IL-36 activity promotes clonal CD4(+) T-cell responses with IL-17A production in generalized pustular psoriasis. J. Investig. Dermatol. 138, 1338–1347 (2018).
Swindell, W. R. et al. RNA-Seq analysis of IL-1B and IL-36 responses in epidermal keratinocytes identifies a shared MyD88-dependent gene signature. Front. Immunol. 9, 80 (2018).
Catapano, M. et al. IL-36 promotes systemic IFN-I responses in severe forms of psoriasis. J. Investig. Dermatol. 140, 816–826.e813 (2020).
Twelves, S. et al. Clinical and genetic differences between pustular psoriasis subtypes. J. Allergy Clin. Immunol. 143, 1021–1026 (2019).
Johnston, A. et al. IL-1 and IL-36 are dominant cytokines in generalized pustular psoriasis. J. Allergy Clin. Immunol. 140, 109–120 (2017).
Marrakchi, S. et al. Interleukin-36-receptor antagonist deficiency and generalized pustular psoriasis. N. Engl. J. Med. 365, 620–628 (2011).
Bachelez, H. et al. Trial of spesolimab for generalized pustular psoriasis. N. Engl. J. Med. 385, 2431–2440 (2021).
Morita, A. et al. Efficacy and safety of subcutaneous spesolimab for the prevention of generalised pustular psoriasis flares (Effisayil 2): an international, multicentre, randomised, placebo-controlled trial. Lancet 402, 1541–1551 (2023).
Henry, C. M. et al. Neutrophil-derived proteases escalate inflammation through activation of IL-36 family cytokines. Cell Rep. 14, 708–722 (2016).
Iznardo, H. & Puig, L. Exploring the role of IL-36 cytokines as a new target in psoriatic disease. Int. J. Mol. Sci. 22, https://doi.org/10.3390/ijms22094344 (2021).
Ma, F. et al. Single-cell and spatial sequencing define processes by which keratinocytes and fibroblasts amplify inflammatory responses in psoriasis. Nat. Commun. 14, 3455 (2023).
Hoegler, K. M., John, A. M., Handler, M. Z. & Schwartz, R. A. Generalized pustular psoriasis: a review and update on treatment. J. Eur. Acad. Dermatol. Venereol. 32, 1645–1651 (2018).
Francis, L. et al. Single-cell analysis of psoriasis resolution demonstrates an inflammatory fibroblast state targeted by IL-23 blockade. Nat. Commun. 15, 913 (2024).
Ma, F. et al. Systems-based identification of the Hippo pathway for promoting fibrotic mesenchymal differentiation in systemic sclerosis. Nat. Commun. 15, 210 (2024).
Stuart, T. et al. Comprehensive integration of single-cell data. Cell 177, 1888–1902.e1821 (2019).
West, A. P. et al. TLR signalling augments macrophage bactericidal activity through mitochondrial ROS. Nature 472, 476–480 (2011).
Yang, L. et al. Hsa_circ_0004287 inhibits macrophage-mediated inflammation in an N(6)-methyladenosine-dependent manner in atopic dermatitis and psoriasis. J. Allergy Clin. Immunol. 149, 2021–2033 (2022).
Trapnell, C. et al. The dynamics and regulators of cell fate decisions are revealed by pseudotemporal ordering of single cells. Nat. Biotechnol. 32, 381–386 (2014).
Sarode, P. et al. Reprogramming of tumor-associated macrophages by targeting β-catenin/FOSL2/ARID5A signaling: a potential treatment of lung cancer. Sci. Adv. 6, eaaz6105 (2020).
Duong, E. et al. Type I interferon activates MHC class I-dressed CD11b(+) conventional dendritic cells to promote protective anti-tumor CD8(+) T cell immunity. Immunity 55, 308–323.e309 (2022).
Walker, J. G. et al. Characterisation of a dendritic cell subset in synovial tissue which strongly expresses Jak/STAT transcription factors from patients with rheumatoid arthritis. Ann. Rheum. Dis. 66, 992–999 (2007).
Chen, J. et al. CREB1-driven CXCR4(hi) neutrophils promote skin inflammation in mouse models and human patients. Nat. Commun. 14, 5894 (2023).
Wigerblad, G. & Kaplan, M. J. Neutrophil extracellular traps in systemic autoimmune and autoinflammatory diseases. Nat. Rev. Immunol. 23, 274–288 (2023).
Brinkmann, V. et al. Neutrophil extracellular traps kill bacteria. Science 303, 1532–1535 (2004).
Oberst, A. et al. Catalytic activity of the caspase-8-FLIP(L) complex inhibits RIPK3-dependent necrosis. Nature 471, 363–367 (2011).
Weinlich, R., Oberst, A., Beere, H. M. & Green, D. R. Necroptosis in development, inflammation and disease. Nat. Rev. Mol. Cell Biol. 18, 127–136 (2017).
Gulati, G. S. et al. Single-cell transcriptional diversity is a hallmark of developmental potential. Science 367, 405–411 (2020).
Haskamp, S. et al. Transcriptomes of MPO-deficient patients with generalized pustular psoriasis reveal expansion of CD4(+) Cytotoxic T cells and an involvement of the complement system. J. Investig. Dermatol. 142, 2149–2158.e2110 (2022).
Hall, J. A. et al. Transcription factor RORα enforces stability of the Th17 cell effector program by binding to a Rorc cis-regulatory element. Immunity 55, 2027–2043.e2029 (2022).
Drummond, R. A. et al. CARD9(+) microglia promote antifungal immunity via IL-1β- and CXCL1-mediated neutrophil recruitment. Nat. Immunol. 20, 559–570 (2019).
Yu, K. et al. Bacterial indole-3-lactic acid affects epithelium-macrophage crosstalk to regulate intestinal homeostasis. Proc. Natl. Acad. Sci. USA 120, e2309032120 (2023).
Hume, D. A. & MacDonald, K. P. Therapeutic applications of macrophage colony-stimulating factor-1 (CSF-1) and antagonists of CSF-1 receptor (CSF-1R) signaling. Blood 119, 1810–1820 (2012).
Guilliams, M., Thierry, G. R., Bonnardel, J. & Bajenoff, M. Establishment and maintenance of the macrophage niche. Immunity 52, 434–451 (2020).
Li, X. et al. The dynamically evolving cell states and ecosystem from benign nevi to melanoma. Clin. Cancer Res. https://doi.org/10.1158/1078-0432.ccr-24-2971 (2025).
Pan, X. et al. Tumour vasculature at single-cell resolution. Nature 632, 429–436 (2024).
Ahmad, F. et al. Emerging role of the IL-36/IL-36R axis in multiple inflammatory skin diseases. J. Investig. Dermatol. 144, 206–224 (2024).
Xu, W. D., Li, R. & Huang, A. F. Role of TL1A in inflammatory autoimmune diseases: a comprehensive review. Front. Immunol. 13, 891328 (2022).
Bamias, G. et al. Expression, localization, and functional activity of TL1A, a novel Th1-polarizing cytokine in inflammatory bowel disease. J. Immunol. 171, 4868–4874 (2003).
Dickson, M. A. et al. Human keratinocytes that express hTERT and also bypass a p16(INK4a)-enforced mechanism that limits life span become immortal yet retain normal growth and differentiation characteristics. Mol. Cell. Biol. 20, 1436–1447 (2000).
Frisch, S. M. Caspase-8: fly or die. Cancer Res. 68, 4491–4493 (2008).
Newton, K. et al. Cleavage of RIPK1 by caspase-8 is crucial for limiting apoptosis and necroptosis. Nature 574, 428–431 (2019).
Lentini, G. et al. Caspase-8 inhibition improves the outcome of bacterial infections in mice by promoting neutrophil activation. Cell Rep. Med. 4, 101098 (2023).
Towne, J. E. et al. Interleukin-36 (IL-36) ligands require processing for full agonist (IL-36α, IL-36β, and IL-36γ) or antagonist (IL-36Ra) activity. J. Biol. Chem. 286, 42594–42602 (2011).
Bayry, J. Immunology: TL1A in the inflammatory network in autoimmune diseases. Nat. Rev. Rheumatol. 6, 67–68 (2010).
Danese, S. et al. Anti-TL1A antibody PF-06480605 safety and efficacy for ulcerative colitis: a phase 2a single-arm Study. Clin. Gastroenterol. Hepatol. 19, 2324–2332.e2326 (2021).
Chinnaiyan, A. M. et al. Signal transduction by DR3, a death domain-containing receptor related to TNFR-1 and CD95. Science274, 990–992 (1996).
Pobezinskaya, Y. L., Choksi, S., Morgan, M. J., Cao, X. & Liu, Z. G. The adaptor protein TRADD is essential for TNF-like ligand 1A/death receptor 3 signaling. J. Immunol. 186, 5212–5216 (2011).
Migone, T. S. et al. TL1A is a TNF-like ligand for DR3 and TR6/DcR3 and functions as a T cell costimulator. Immunity 16, 479–492 (2002).
Bamias, G. et al. Upregulation and nuclear localization of TNF-like cytokine 1A (TL1A) and its receptors DR3 and DcR3 in psoriatic skin lesions. Exp. Dermatol. 20, 725–731 (2011).
Bamias, G., Menghini, P., Pizarro, T. T. & Cominelli, F. Targeting TL1A and DR3: the new frontier of anti-cytokine therapy in IBD. Gut 74, 652–668 (2025).
McGuire, J. & Arnesen, S. Control of keratinocyte division in vitro. J. Investig. Dermatol. 59, 84–90 (1972).
Pulido-Pérez, A. & Bergon-Sendin, M. Sweet’s syndrome. N. Engl. J. Med. 382, 1543 (2020).
Sugiura, K. The genetic background of generalized pustular psoriasis: IL36RN mutations and CARD14 gain-of-function variants. J. Dermatol. Sci. 74, 187–192 (2014).
Butler, A., Hoffman, P., Smibert, P., Papalexi, E. & Satija, R. Integrating single-cell transcriptomic data across different conditions, technologies, and species. Nat. Biotechnol. 36, 411–420 (2018).
Sica, A. & Mantovani, A. Macrophage plasticity and polarization: in vivo veritas. J. Clin. Investig. 122, 787–795 (2012).
Subramanian, A. et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc. Natl. Acad. Sci. USA 102, 15545–15550 (2005).
Hänzelmann, S., Castelo, R. & Guinney, J. GSVA: gene set variation analysis for microarray and RNA-seq data. BMC Bioinform. 14, 7 (2013).
Corridoni, D. et al. Single-cell atlas of colonic CD8(+) T cells in ulcerative colitis. Nat. Med. 26, 1480–1490 (2020).
Billi, A. C. et al. Nonlesional lupus skin contributes to inflammatory education of myeloid cells and primes for cutaneous inflammation. Sci. Transl. Med. 14, eabn2263 (2022).
Love, M. I., Huber, W. & Anders, S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genom. Biol. 15, 550 (2014).
Jin, S. et al. Inference and analysis of cell–cell communication using CellChat. Nat. Commun. 12, 1088 (2021).
Browaeys, R., Saelens, W. & Saeys, Y. NicheNet: modeling intercellular communication by linking ligands to target genes. Nat. Methods 17, 159–162 (2020).
Janesick, A. et al. High-resolution mapping of the tumor microenvironment using integrated single-cell, spatial and in situ analysis. Nat. Commun. 14, 8353 (2023).
Acknowledgements
This work was supported by a non-restricted research grant from Boehringer Ingelheim (J.E.G.), the National Institutes of Health (K01AR072129 and R01AR080662 to L.C.T.; 1P30AR075043 to L.C.T., M.T.P., and J.E.G.; UC2 AR081033 to L.C.T. and J.E.G.). Sinocare Diabetes Foundation Scholarship for the Michigan-Xiangya MD/PhD Dual Degree Program (awarded to R.J.).
Author information
Authors and Affiliations
Contributions
Conceptualization: R.J. and J.E.G.; Methodology: R.J., J.K., J.F., J.W., R.B., M.K.S., S.S., and J.E.G.; Imaging: X.X. and R.J.; Investigation: R.J., J.K., J.F., X.X., J.W., M.K.S., R.B., T.D., A.C., C.C., O.P., J.E.R., H.Z., J.M.K., P.W.H., H.B., L.C.T., S.S., and A.C.B.; Writing: R.J. and J.E.G.; Funding Acquisition: L.C.T. and J.E.G. Supervision: H.B., X.C., and J.E.G.
Corresponding authors
Ethics declarations
Competing interests
This was an independent, investigator-initiated study supported by Boehringer Ingelheim Pharmaceuticals, Inc. (BIPI). BIPI had no role in the design, analysis or interpretation of the results in this study. BIPI was given the opportunity to review the manuscript for medical and scientific accuracy as it relates to BIPI substances, as well as intellectual property considerations. The remaining authors declare no competing interests.
Peer review
Peer review information
Nature Communications thanks Mayumi Komine, Shigetoshi Sano and the other anonymous reviewer(s) for their contribution to the peer review of this work. A peer review file is available.
Additional information
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary information
Source data
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.
About this article
Cite this article
Jiang, R., Kirma, J., Fox, J. et al. Dynamic neutrophil-keratinocyte communication network centered on IL-36/TNFSF15 responses characterizes inflammatory responses in generalized pustular psoriasis. Nat Commun (2025). https://doi.org/10.1038/s41467-025-67917-9
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41467-025-67917-9
