{
  "abstract": "Human monocytes are classified as classical (CD14++CD16−), intermediate (CD14++CD16+), and non-classical (CD14+CD16++) subsets, but CD16 stability during ex vivo assays remains uncertain. Here, we examined whether standard PBMC handling alters CD16-based subset identity and whether ADAM17 inhibition can prevent this artifact. In isolated PBMCs, intermediate and non-classical monocytes rapidly lost surface CD16 during culture at 37 °C, with or without LPS stimulation, whereas CD16 expression remained stable at 4 °C and in unseparated whole blood. Pre- versus post-incubation labeling and sorted-cell experiments indicated that CD16 loss was not primarily due to receptor internalization or cell death, but instead produced a pseudo-classical phenotype. Public scRNA-seq analyses showed enriched ADAM17 expression in monocyte/macrophage populations across blood, liver, and kidney datasets. Pharmacological ADAM17 inhibition with TMI-005 or TAPI-1 preserved CD16 expression during LPS stimulation without broadly impairing monocyte cytokine production. We therefore propose an ADAM17 inhibitor-based protocol that combines surface staining, stimulation, and intracellular cytokine profiling to preserve monocyte subset identity during functional assays. These findings identify a practical strategy to reduce CD16-dependent monocyte subset misclassification in ex vivo immunological studies.",
  "abstract_shown": true,
  "affiliations": [
    "Department of Bio-Medical Science, GAIHST, Gachon University",
    "Department of Microbiology, Gachon University College of Medicine",
    "Division of Gastroenterology and Hepatology, Department of Internal Medicine, Gachon University Gil Medical Center, Gachon University College of Medicine"
  ],
  "article_type": "research article",
  "arxiv": null,
  "author_total": 3,
  "authors": [
    {
      "affs": [
        0,
        1
      ],
      "name": "Soyeon Kwon"
    },
    {
      "affs": [
        2
      ],
      "name": "Seung Kak Shin"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Seungwon Ryu"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/3yznbxffm969b.bib",
  "cited_by": 0,
  "doi": "10.3389/fimmu.2026.1886931",
  "fulltext_url": "https://europepmc.org/articles/PMC13710362",
  "id": "3yznbxffm969b",
  "issns": [
    "1664-3224"
  ],
  "landing_url": "https://www.frontiersin.org/articles/10.3389/fimmu.2026.1886931/full",
  "language": "eng",
  "licence": "CC-BY-4.0",
  "licence_label": "CC BY 4.0",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "heading",
      "Original Research"
    ],
    [
      "openalex",
      "2403|Immune cells in cancer"
    ]
  ],
  "openalex": "W7221608485",
  "pdf_url": "https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2026.1886931/pdf",
  "pmcid": "PMC13710362",
  "pmid": "42948125",
  "published": "2026-10-13",
  "publisher": "Frontiers Media SA",
  "ris": "https://research.draconitic.com/paper/3yznbxffm969b.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:19:15Z",
      "native_id": "10.3389/fimmu.2026.1886931",
      "source": "crossref",
      "url": "https://www.frontiersin.org/articles/10.3389/fimmu.2026.1886931/full"
    },
    {
      "first_seen": "2026-10-31T00:20:34Z",
      "native_id": "MED:42948125",
      "source": "europepmc",
      "url": "https://europepmc.org/article/MED/42948125"
    },
    {
      "first_seen": "2026-10-31T12:21:21Z",
      "native_id": "PMC13710362",
      "source": "pmc",
      "url": "https://doi.org/10.3389/fimmu.2026.1886931"
    }
  ],
  "subjects": [
    {
      "field": "biology",
      "origin": "openalex:2403|Immune cells in cancer",
      "subfield": "immunology"
    }
  ],
  "title": "ADAM17-mediated CD16 shedding drives monocyte subset misclassification in ex vivo immunological assays",
  "title_text": "ADAM17-mediated CD16 shedding drives monocyte subset misclassification in ex vivo immunological assays",
  "type": "peer-reviewed",
  "updated_at": "2026-10-31T12:21:21Z",
  "url": "https://research.draconitic.com/paper/3yznbxffm969b",
  "venue": "Frontiers in Immunology",
  "versions": []
}