Flow Cytometry Assessment of Lymphocyte Populations Infiltrating Liver Tumors.

Autor:	Pérez-Lanzón M; Centre de Recherche des Cordeliers, Equipe labellisée par la Ligue contre le cancer, Inserm U1138, Université Paris Cité, Sorbonne Université, Institut Universitaire de France, Paris, France.; Metabolomics and Cell Biology Platforms, UMS AMMICa, Gustave Roussy, Villejuif, France., Plantureux C; Centre de Recherche des Cordeliers, Equipe labellisée par la Ligue contre le cancer, Inserm U1138, Université Paris Cité, Sorbonne Université, Institut Universitaire de France, Paris, France.; Metabolomics and Cell Biology Platforms, UMS AMMICa, Gustave Roussy, Villejuif, France.; Faculté de Médecine, Université Paris-Saclay, Kremlin-Bicêtre, France., Paillet J; Centre de Recherche des Cordeliers, Equipe labellisée par la Ligue contre le cancer, Inserm U1138, Université Paris Cité, Sorbonne Université, Institut Universitaire de France, Paris, France.; Metabolomics and Cell Biology Platforms, UMS AMMICa, Gustave Roussy, Villejuif, France.; Laboratory of Human Lymphohematopoieisis, Imagine Institute, INSERM UMR 1163, Université Paris Cité, Paris, France.; Smart Immune, Paris, France., Sotty J; Sorbonne Université, Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Recherche de Saint Antoine (CRSA), Paris, France., Soussan P; Sorbonne Université, Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Recherche de Saint Antoine (CRSA), Paris, France.; Assistance Publique - Hôpitaux de Paris (AP-HP). Sorbonne Université, Département de Virologie, GHU Paris-Est, Paris, France., Kroemer G; Centre de Recherche des Cordeliers, Equipe labellisée par la Ligue contre le cancer, Inserm U1138, Université Paris Cité, Sorbonne Université, Institut Universitaire de France, Paris, France.; Metabolomics and Cell Biology Platforms, UMS AMMICa, Gustave Roussy, Villejuif, France.; Institut du Cancer Paris CARPEM, Department of Biology, Hôpital Européen Georges Pompidou, AP-HP, Paris, France., Maiuri MC; Centre de Recherche des Cordeliers, Equipe labellisée par la Ligue contre le cancer, Inserm U1138, Université Paris Cité, Sorbonne Université, Institut Universitaire de France, Paris, France.; Metabolomics and Cell Biology Platforms, UMS AMMICa, Gustave Roussy, Villejuif, France.; Department of Molecular Medicine and Medical Biotechnologies, University of Napoli Federico II, Naples, Italy., Pol J; Centre de Recherche des Cordeliers, Equipe labellisée par la Ligue contre le cancer, Inserm U1138, Université Paris Cité, Sorbonne Université, Institut Universitaire de France, Paris, France.; Metabolomics and Cell Biology Platforms, UMS AMMICa, Gustave Roussy, Villejuif, France.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2024; Vol. 2769, pp. 129-141.
DOI:	10.1007/978-1-0716-3694-7_10
Abstrakt:	Tissue-resident and recruited immune cells are essential mediators of natural and therapy-induced immunosurveillance of liver neoplasia. This idea has been recently reinforced by the clinical approval of immune checkpoint inhibitors for the immunotherapy of hepatocellular carcinoma and cholangiocarcinoma. Such research progress relies on the in-depth characterization of the immune populations that are present in pre-neoplastic and neoplastic hepatic lesions. A convenient technology for advancing along this path is high-dimensional cytometry.In this chapter, we present a protocol to assess the subtype and differentiation state of hepatic lymphocyte populations by multicolor immunofluorescence staining and flow cytometry. We detail the steps required for viability assessment and immune cell phenotyping of single-cell suspensions of liver cells by means of surface and intracellular staining of more than a dozen markers of interest. This protocol does not require prior removal of debris and dead cells and allows to process multiple samples in parallel. The procedure includes the use of a fixative-resistant viability dye that allows cell fixation and permeabilization after cell surface staining and before intracellular staining and data acquisition on a flow cytometer. Moreover, we provide a panel of fluorochrome-labeled antibodies designed for the characterization of lymphocytic subsets that can be adapted to distinct experimental settings. Finally, we present an overview of the post-staining pipeline, including data acquisition on a flow cytometer and tools for post-acquisition analyses. (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze:	MEDLINE
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