{
  "abstract": "Invariant natural killer T (iNKT) cells can limit graft-versus-host disease (GVHD) after hematopoietic stem cell transplantation (HSCT), but their scarcity in peripheral blood limitstherapeutic development. Current clinical-grade human iNKT expansion protocols mainly rely on IL-2, require prior iNKT-cell sorting, last 6-8 weeks, and predominantly expand CD4+ iNKT cells, whereas human CD4- iNKT cells are more strongly associated with GVHD control in patients and uniquely regulate antigen-presenting cells and T-cell activation. We developed a scalable culture system to preferentially expand human CD4- iNKT cells directly from total peripheral blood mononuclear cells (PBMCs) using alpha galactosylceramide (-GalCer) and optimized cytokine conditions. IL-15 was the most effective cytokine. The optimized 14-day protocol generated a mean of 3.8x107 iNKT cells from 2x107 PBMCs, including 74% CD4- iNKT cells. Single-cell transcriptomic profiling identified eight major iNKT subsets, differentiation trajectories during expansion, and distinct IL-2- versus IL-15-associated transcriptional programs. IL-15-expanded iNKT cells induced apoptosis of monocyte-derived dendritic and leukemic cells in vitro, controlled xeno-GVHD, and preserved graft-versus-leukemia (GVL) activity in preclinical mouse models. This platform enables reproducible production of human CD4- iNKT cells at clinically relevant scale and position IL-15-expanded iNKT cells as a compelling immunotherapy candidate for allo-HSCT.",
  "abstract_shown": true,
  "affiliations": [
    "IMoPA Universite de Lorraine CNRS"
  ],
  "article_type": "New results",
  "arxiv": null,
  "author_total": 9,
  "authors": [
    {
      "name": "J. Brouard"
    },
    {
      "name": "C. Caraiman"
    },
    {
      "name": "G. FIEVET"
    },
    {
      "name": "L. Monchablon"
    },
    {
      "name": "T. Coman"
    },
    {
      "name": "S. Hergalant"
    },
    {
      "name": "S. Pagliuca"
    },
    {
      "name": "D. MOULIN"
    },
    {
      "affs": [
        0
      ],
      "name": "M. T. Rubio"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/57zs0dnen21fq.bib",
  "cited_by": 0,
  "doi": "10.64898/2026.09.11.750594",
  "fulltext_url": "https://www.biorxiv.org/content/10.64898/2026.09.11.750594v1.full",
  "id": "57zs0dnen21fq",
  "issns": [],
  "landing_url": "https://www.biorxiv.org/content/10.64898/2026.09.11.750594v1",
  "language": null,
  "licence": "CC-BY-NC-ND",
  "licence_label": "CC BY-NC-ND",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by-nc-nd/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "biorxiv",
      "immunology"
    ],
    [
      "openalex",
      "2720|Hematopoietic Stem Cell Transplantation"
    ]
  ],
  "openalex": "W7213446342",
  "pdf_url": "https://www.biorxiv.org/content/10.64898/2026.09.11.750594v1.full.pdf",
  "pmcid": null,
  "pmid": null,
  "published": "2026-09-11",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/57zs0dnen21fq.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:22:43Z",
      "native_id": "10.64898/2026.09.11.750594",
      "source": "biorxiv",
      "url": "https://www.biorxiv.org/content/10.64898/2026.09.11.750594v1"
    }
  ],
  "subjects": [
    {
      "field": "biology",
      "origin": "biorxiv:immunology",
      "subfield": "immunology"
    }
  ],
  "title": "Scalable expansion of human iNKT cells: single-cell profiling and in vivo control of GvHD with preserved GvL activity",
  "title_text": "Scalable expansion of human iNKT cells: single-cell profiling and in vivo control of GvHD with preserved GvL activity",
  "type": "preprint",
  "updated_at": "2026-10-30T12:22:43Z",
  "url": "https://research.draconitic.com/paper/57zs0dnen21fq",
  "venue": "bioRxiv",
  "versions": []
}