{
  "abstract": "Background Intrahepatic cholangiocarcinoma (iCCA) is a highly aggressive biliary tract cancer with a poor prognosis and a complex tumour microenvironment (TME) that remains poorly understood.Objective This study aimed to investigate the phenotypic and molecular characteristics of B lymphocytes, their interactions with the TME and their prognostic implications.Design B-cell compartments in the tumour, peritumour, and peripheral blood of iCCA patients were analysed using multimodal single-cell technologies. The B-cell interactome with the iCCA TME was explored in silico, and ex vivo assays assessed the impact of interactions with cancer-associated fibroblasts (CAFs) and tumour cells on B-cell biology. B-cell modulation during chemoimmunotherapy in advanced iCCA was also evaluated.Results B cells were enriched in adjacent tumour-free tissues and formed mature tertiary lymphoid structures (TLS), correlating with better prognosis. Conversely, tumour-infiltrating B cells were scarce, immature and displayed reduced effector function with increased immunosuppressive features. Coculture with tumour cells or CAFs impaired B-cell differentiation and function, including downregulation of BAFFR in peripheral B cells. IL-6 and TGF-β emerged as major drivers of B-cell dysfunction; dual blockade restored B-cell activation and differentiation. Elevated frequencies of circulating BAFFR + B cells and hyperexpanded clonotypes were linked to improved chemoimmunotherapy response.Conclusions iCCA is characterised by a profoundly immunosuppressive TME that impairs B-cell function through soluble factors and cellular interactions. Our findings identify B cells as biomarkers and therapeutic targets, supporting strategies to restore B-cell function and promote mature TLS to enhance immunotherapy responsiveness in iCCA.",
  "authors": [
    {
      "affiliations": [
        "Hepatobiliary Immunopathology Lab, Department of Gastroenterology, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Giulia Milardi"
    },
    {
      "affiliations": [
        "Hepatobiliary Immunopathology Lab, Department of Gastroenterology, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Barbara Franceschini"
    },
    {
      "affiliations": [
        "Flow Cytometry Core, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Chiara Camisaschi"
    },
    {
      "affiliations": [
        "Institute of Genetic and Biomedical Research, UoS Milan, National Research Council, Rozzano, (MI), Italy",
        "Laboratory of Translational Immunology, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Simone Puccio"
    },
    {
      "affiliations": [
        "Division of Hepatobiliary and General Surgery, Department of Surgery, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy",
        "Department of Biomedical Sciences, Humanitas University, Pieve Emanuele (MI), Italy"
      ],
      "name": "Guido Costa"
    },
    {
      "affiliations": [
        "Hepatobiliary Immunopathology Lab, Department of Gastroenterology, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Cristiana Soldani"
    },
    {
      "affiliations": [
        "Clinical Bioinformatics Unit, IRCCS Istituto Giannina Gaslini, Genoa, Italy"
      ],
      "name": "Paolo Uva"
    },
    {
      "affiliations": [
        "Clinical Bioinformatics Unit, IRCCS Istituto Giannina Gaslini, Genoa, Italy"
      ],
      "name": "Davide Cangelosi"
    },
    {
      "affiliations": [
        "Bioinformatics Unit, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Roberta Carriero"
    },
    {
      "affiliations": [
        "Bioinformatics Unit, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Luca Lambroia"
    },
    {
      "affiliations": [
        "Hepatobiliary Immunopathology Lab, Department of Gastroenterology, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy",
        "Department of Biomedical Sciences, Humanitas University, Pieve Emanuele (MI), Italy"
      ],
      "name": "Antonella Cammarota"
    },
    {
      "affiliations": [
        "Hepatobiliary Immunopathology Lab, Department of Gastroenterology, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Giulio Lodetti-Zangrandi"
    },
    {
      "affiliations": [
        "Laboratory of Translational Immunology, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Ines Malenica"
    },
    {
      "affiliations": [
        "Department of Biomedical Sciences, Humanitas University, Pieve Emanuele (MI), Italy",
        "Unit of Multiscale and Nanostructural Imaging, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Marco Erreni"
    },
    {
      "affiliations": [
        "Flow Cytometry Core, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Ilaria Montali"
    },
    {
      "affiliations": [
        "Department of Experimental and Clinical Medicine, Università degli Studi di Firenze, Florence, Italy"
      ],
      "name": "Tiziano Lottini"
    },
    {
      "affiliations": [
        "Department of Experimental and Clinical Medicine, Università degli Studi di Firenze, Florence, Italy"
      ],
      "name": "Chiara Raggi"
    },
    {
      "affiliations": [
        "Bioinformatics Unit, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Paolo Kunderfranco"
    },
    {
      "affiliations": [
        "Hepatobiliary Immunopathology Lab, Department of Gastroenterology, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Michela Anna Polidoro"
    },
    {
      "affiliations": [
        "Department of Biomedical Sciences, Humanitas University, Pieve Emanuele (MI), Italy",
        "Division of Internal Medicine and Hepatology, Department of Gastroenterology, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Alessio Aghemo"
    },
    {
      "affiliations": [
        "Department of Biomedical Sciences, Humanitas University, Pieve Emanuele (MI), Italy",
        "Medical Oncology and Hematology Unit, Humanitas Cancer Center, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Rita Balsano"
    },
    {
      "affiliations": [
        "Medical Oncology and Hematology Unit, Humanitas Cancer Center, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Tiziana Pressiani"
    },
    {
      "affiliations": [
        "Department of Biomedical Sciences, Humanitas University, Pieve Emanuele (MI), Italy",
        "Pathology Unit, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Salvatore Piscuoglio"
    },
    {
      "affiliations": [
        "Department of Biomedical Sciences, Humanitas University, Pieve Emanuele (MI), Italy",
        "Pathology Unit, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Luca Di Tommaso"
    },
    {
      "affiliations": [
        "Division of Hepatobiliary and General Surgery, Department of Surgery, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy",
        "Department of Biomedical Sciences, Humanitas University, Pieve Emanuele (MI), Italy"
      ],
      "name": "Guido Torzilli"
    },
    {
      "affiliations": [
        "Department of Biomedical Sciences, Humanitas University, Pieve Emanuele (MI), Italy",
        "Medical Oncology and Hematology Unit, Humanitas Cancer Center, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Lorenza Rimassa"
    },
    {
      "affiliations": [
        "Laboratory of Translational Immunology, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Enrico Lugli"
    },
    {
      "affiliations": [
        "Department of Medical Biotechnologies and Translational Medicine, Università degli Studi di Milano, Milan, Italy",
        "Immunity and Immunodeficiency Lab, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Barbara Cassani"
    },
    {
      "affiliations": [
        "Department of Biomedical Sciences, Humanitas University, Pieve Emanuele (MI), Italy",
        "Division of Internal Medicine and Hepatology, Department of Gastroenterology, IRCCS Humanitas Research Hospital, Rozzano (MI), Italy"
      ],
      "name": "Ana Lleo"
    }
  ],
  "title": "Immunosuppressive contribution of tumour-infiltrating B cells in human intrahepatic cholangiocarcinoma and their role in chemoimmunotherapy outcome",
  "uid": "6d83e5fb-bb7c-59db-8c24-3887b4235e54"
}