Single-cell analysis reveals cell death as driver of NLRP3-mediated secretion of IL-1β in human monocytes

  • Nat Immunol. 2025 Dec;26(12):2148-2158. doi: 10.1038/s41590-025-02319-z.
Lieselotte Vande Walle  #  1 ,  Kentaro Kato  #  2 ,  Mai Yamagishi  3 ,  Takashi Kamatani  4  5 ,  Alex Vervaeke  1 ,  Rosa Martín-Pérez  6 ,  Masaki Shimizu  7 ,  Takumi Takizawa  8 ,  Junko Takita  2 ,  Ryuta Nishikomori  9 ,  Osamu Ohara  10 ,  Kazushi Izawa  11 ,  Yoshitaka Shirasaki  12 ,  Mohamed Lamkanfi  13
Affiliations
  • 1. Laboratory of Medical Immunology, Department of Internal Medicine and Paediatrics, Ghent University, Ghent, Belgium.
  • 2. Department of Pediatrics, Kyoto University Graduate School of Medicine, Kyoto, Japan.
  • 3. Live Cell Diagnosis, Ltd, Saitama, Japan.
  • 4. Department of AI Technology Development, M&D Data Science Center, Institute of Integrated Research, Institute of Science Tokyo, Tokyo, Japan.
  • 5. Department of Precision Cancer Medicine, Center for Innovative Cancer Treatment, Institute of Science Tokyo Hospital, Tokyo, Japan.
  • 6. Janssen Interventional Oncology, Beerse, Belgium.
  • 7. Department of Pediatrics, Perinatal and Maternal Medicine, Institute of Science Tokyo, Tokyo, Japan.
  • 8. Department of Pediatrics, Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan.
  • 9. Department of Pediatrics and Child Health, Kurume University School of Medicine, Kurume, Japan.
  • 10. KAZUSA DNA Research Institute, Kisarazu, Chiba, Japan.
  • 11. Department of Pediatrics, Kyoto University Graduate School of Medicine, Kyoto, Japan. [email protected].
  • 12. Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, Japan. [email protected].
  • 13. Laboratory of Medical Immunology, Department of Internal Medicine and Paediatrics, Ghent University, Ghent, Belgium. [email protected].
  • # Contributed equally.
PMID: 41219490 DOI: 10.1038/s41590-025-02319-z
Abstract

Interleukin-1β (IL-1β) is a key proinflammatory cytokine with critical roles in infections and inflammatory diseases, yet the mechanisms regulating its release from human monocytes remain unclear. Here we used a suite of single-cell approaches, including integrated live-cell imaging of secretion and cell fate, flow cytometry and high-content imaging, to investigate IL-1β secretion dynamics in lipopolysaccharide-stimulated primary human peripheral blood CD14+ monocytes. We found marked heterogeneity: a large fraction of cells remained viable and contributed negligibly to IL-1β secretion, challenging established models. Instead, a small subset (5-10%) undergoing canonical NLRP3 inflammasome activation and GSDMD-dependent Pyroptosis produced the majority of secreted IL-1β, with a smaller contribution from apoptotic cells transitioning to secondary necrosis. Single-cell profiling of CD14+ monocytes from patients with cryopyrin-associated periodic syndrome confirmed lytic cell death as the driver of pathological IL-1β release. These findings redefine IL-1β as a damage-associated molecular pattern, secreted predominantly by dying monocytes.

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