title: Immune cell dysfunctions in breast cancer patients detected through whole blood multi-parametric flow cytometry assay.
abstract: Monitoring functional competence of immune cell populations in clinical routine represents a major challenge. We developed a whole-blood assay to monitor functional competence of peripheral innate immune cells including NK cells, dendritic and monocyte cell subsets through their ability to produce specific cytokines after short-term stimulation, detected through intra-cytoplasmic staining and multi-parametric flow-cytometry. A PMA/ionomycin T cell activation assay complemented this analysis. Comparing cohorts of healthy women and breast cancer (BC) patients at different stages, we identified significant functional alteration of circulating immune cells during BC progression prior to initiation of treatment. Of upmost importance, as early as the localized primary tumor (PT) stage, we observed functional alterations in several innate immune populations and T cells i.e. (i) reduced TNFα production by BDCA-1+DC and non-classical monocytes in response to Type-I IFN, (ii) a strong drop in IFNγ production by NK cells in response to either Type-I IFN or TLR7/8 ligand, and (iii) a coordinated impairment of cytokine (IL-2, IFNγ, IL-21) production by T cell subpopulations. Overall, these alterations are further accentuated according to the stage of the disease in first-line metastatic patients. Finally, whereas we did not detect functional modification of DC subsets in response to TLR7/8 ligand, we highlighted increased IL-12p40 production by monocytes specifically at first relapse (FR). Our results reinforce the importance of monitoring both innate and adaptive immunity to better evaluate dysfunctions in cancer patients and suggest that our whole-blood assay will be useful to monitor response to treatment, particularly for immunotherapeutic strategies.
authors.affiliation: Innovation in Immuno-monitoring and Immunotherapy Platform (PI3), Léon Bérard Cancer Center , Lyon, France.
authors.forename: E
authors.initials: E
authors.lastname: Verronèse
authors.affiliation: Innovation in Immuno-monitoring and Immunotherapy Platform (PI3), Léon Bérard Cancer Center , Lyon, France.
authors.forename: A
authors.initials: A
authors.lastname: Delgado
authors.affiliation: Team 11, INSERM U1052/CNRS UMR5286, Cancer Research Center of Lyon, Lyon, France; Université de Lyon, Lyon, France; Université Lyon 1, ISPB, Lyon, France.
authors.forename: J
authors.initials: J
authors.lastname: Valladeau-Guilemond
authors.affiliation: DRCI department, Léon Bérard Cancer Center , Lyon, France.
authors.forename: G
authors.initials: G
authors.lastname: Garin
authors.affiliation: DRCI department, Léon Bérard Cancer Center , Lyon, France.
authors.forename: S
authors.initials: S
authors.lastname: Guillemaut
authors.affiliation: Department of Medical Oncology, Léon Bérard Cancer Center , Lyon, France.
authors.forename: O
authors.initials: O
authors.lastname: Tredan
authors.affiliation: Department of Medical Oncology, Léon Bérard Cancer Center , Lyon, France.
authors.forename: I
authors.initials: I
authors.lastname: Ray-Coquard
authors.affiliation: Team 11, INSERM U1052/CNRS UMR5286, Cancer Research Center of Lyon, Lyon, France; Université de Lyon, Lyon, France; Université Lyon 1, ISPB, Lyon, France; Department of Medical Oncology, Léon Bérard Cancer Center, Lyon, France.
authors.forename: T
authors.initials: T
authors.lastname: Bachelot
authors.affiliation: Innovation in Immuno-monitoring and Immunotherapy Platform (PI3), Léon Bérard Cancer Center , Lyon, France.
authors.forename: A
authors.initials: A
authors.lastname: N'Kodia
authors.affiliation: Innovation in Immuno-monitoring and Immunotherapy Platform (PI3), Léon Bérard Cancer Center , Lyon, France.
authors.forename: C
authors.initials: C
authors.lastname: Bardin-Dit-Courageot
authors.affiliation: Innovation in Immuno-monitoring and Immunotherapy Platform (PI3), Léon Bérard Cancer Center , Lyon, France.
authors.forename: C
authors.initials: C
authors.lastname: Rigal
authors.affiliation: DRCI department, Léon Bérard Cancer Center , Lyon, France.
authors.forename: D
authors.initials: D
authors.lastname: Pérol
authors.affiliation: Innovation in Immuno-monitoring and Immunotherapy Platform (PI3), Léon Bérard Cancer Center, Lyon, France; Team 11, INSERM U1052/CNRS UMR5286, Cancer Research Center of Lyon, Lyon, France; Université de Lyon, Lyon, France; Université Lyon 1, ISPB, Lyon, France.
authors.forename: C
authors.initials: C
authors.lastname: Caux
authors.affiliation: Innovation in Immuno-monitoring and Immunotherapy Platform (PI3), Léon Bérard Cancer Center, Lyon, France; Team 11, INSERM U1052/CNRS UMR5286, Cancer Research Center of Lyon, Lyon, France; Université de Lyon, Lyon, France; Université Lyon 1, ISPB, Lyon, France.
authors.forename: C
authors.initials: C
authors.lastname: Ménétrier-Caux
keywords: Breast cancer; immune system; monitoring; multi-parametric flow cytometry; whole blood assay