{
  "abstract": "Background\n\nDirect, cell-resolved quantification of NLRP3 inflammasome activation in clinical patients has not been feasible. Previous studies have relied on indirect surrogates such as downstream cytokines, which are influenced by inflammasome-independent pathways and systemic clearance kinetics, potentially explaining the inconsistent prognostic findings reported in the literature. We applied a proximity ligation assay (PLA) to directly visualise NLRP3-caspase-1 complex formation in peripheral blood mononuclear cells of septic patients, providing a cell-specific, in situ readout of inflammasome assembly at the single-cell level.Methods\n\nIn this prospective, multicentre proof-of-concept pilot study, 78 patients fulfilling Sepsis-3 criteria were enrolled across three university hospitals. The PLA was validated in THP-1-derived macrophages against established readouts of canonical inflammasome activation prior to clinical application. Patients were stratified into low (n = 28), intermediate (n = 23), and high (n = 27) NLRP3 inflammasome activity groups.Results\n\nMedian age was 64 years (IQR: 56-78), 63% were male, and median Sequential Organ Failure Assessment score was 8 (IQR: 4-12). Baseline characteristics, disease severity scores, and routine inflammatory biomarkers did not differ across activity groups. Kaplan-Meier analysis revealed a non-linear pattern of 30-day survival, with the most favourable outcomes in the intermediate activity group. Both low and high activity were associated with increased mortality (log-rank p = 0.003). In adjusted Cox regression, low inflammasome activity (HR 7.88, 95% CI 1.79-34.65; p = 0.006) and high inflammasome activity (HR 4.35, 95% CI 1.01-19.88; p = 0.048) remained associated with 30-day mortality. Modelling PLA activity as a continuous variable using restricted cubic splines confirmed a significant non-linear association with 30-day mortality, independent of the tertile-based categorisation (p for non-linearity = 0.015).Conclusions\n\nThis proof-of-concept pilot study demonstrates that direct in situ quantification of NLRP3-caspase-1 complex assembly is feasible in septic patients and reveals a non-linear association with 30-day survival. These findings indicate that PLA-based profiling captures immune heterogeneity beyond routine inflammatory biomarkers. Larger independent cohorts are needed to validate these findings before clinical application or patient stratification can be considered.",
  "abstract_shown": true,
  "affiliations": [
    "Ruhr-Universität Bochum; Klinik für Anästhesiologie, Intensivmedizin und Schmerztherapie; Knappschaft Kliniken - Universitätsklinikum Bochum, In der Schornau 23-25, Bochum, 44892, Germany.",
    "Klinik für Anästhesiologie und Operative Intensivmedizin, Universitätsklinikum Bonn, Bonn, 53127, Germany.",
    "Department of Anesthesiology, Surgical Intensive Care, Emergency and Pain Medicine, Klinikum Herford, Ruhr University Bochum, Herford, 32049, Germany.",
    "Ruhr-Universität Bochum; Klinik für Anästhesiologie, Intensivmedizin und Schmerztherapie; Knappschaft Kliniken - Universitätsklinikum Bochum, In der Schornau 23-25, Bochum, 44892, Germany. Tim.Rahmel@ruhr-uni-bochum.de."
  ],
  "article_type": "research article",
  "arxiv": null,
  "author_total": 12,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Fabian Mehl"
    },
    {
      "affs": [
        0
      ],
      "name": "Willy J Chedjou"
    },
    {
      "affs": [
        0
      ],
      "name": "Andrea Witowski"
    },
    {
      "affs": [
        0
      ],
      "name": "Katharina Rump"
    },
    {
      "affs": [
        0
      ],
      "name": "Birte Dyck"
    },
    {
      "affs": [
        0
      ],
      "name": "Lars Bergmann"
    },
    {
      "affs": [
        1
      ],
      "name": "Christian Putensen"
    },
    {
      "affs": [
        2
      ],
      "name": "Dietrich Henzler"
    },
    {
      "affs": [
        0
      ],
      "name": "Michael Adamzik"
    },
    {
      "affs": [
        0
      ],
      "name": "Björn Koos"
    },
    {
      "affs": [
        0
      ],
      "name": "Patrick Thon"
    },
    {
      "affs": [
        3
      ],
      "name": "Tim Rahmel",
      "orcid": "0000-0002-7039-6288"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/dwy1p4h9yq3hj.bib",
  "cited_by": 0,
  "doi": "10.1186/s40635-026-00992-4",
  "fulltext_url": "https://europepmc.org/articles/PMC13719975",
  "id": "dwy1p4h9yq3hj",
  "issns": [
    "2197-425X"
  ],
  "landing_url": "https://europepmc.org/article/MED/42955163",
  "language": "eng",
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "heading",
      "Research Articles"
    ],
    [
      "openalex",
      "1312|Inflammasome and immune disorders"
    ]
  ],
  "openalex": "W7224187938",
  "pdf_url": "https://europepmc.org/articles/PMC13719975?pdf=render",
  "pmcid": "PMC13719975",
  "pmid": "42955163",
  "published": "2026-10-29",
  "publisher": "Springer",
  "ris": "https://research.draconitic.com/paper/dwy1p4h9yq3hj.ris",
  "sources": [
    {
      "first_seen": "2026-11-01T00:20:36Z",
      "native_id": "MED:42955163",
      "source": "europepmc",
      "url": "https://europepmc.org/article/MED/42955163"
    },
    {
      "first_seen": "2026-11-02T12:23:50Z",
      "native_id": "PMC13719975",
      "source": "pmc",
      "url": "https://doi.org/10.1186/s40635-026-00992-4"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|Inflammasome and immune disorders",
      "subfield": "molecular-biology"
    }
  ],
  "title": "Quantification of NLRP3-caspase-1 complex assembly reveals a non-linear association with 30-day mortality in sepsis: a proof-of-concept pilot study.",
  "title_text": "Quantification of NLRP3-caspase-1 complex assembly reveals a non-linear association with 30-day mortality in sepsis: a proof-of-concept pilot study.",
  "type": "peer-reviewed",
  "updated_at": "2026-11-03T12:20:56Z",
  "url": "https://research.draconitic.com/paper/dwy1p4h9yq3hj",
  "venue": "Intensive care medicine experimental",
  "versions": []
}