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Published on: March 5, 2019
Immune cell dysfunctions in breast cancer patients detected through whole blood multi-parametric flow cytometry assay
E Verronèse1, A Delgado1, J Valladeau-Guilemond2
1Innovation in Immuno-monitoring and Immunotherapy Platform (PI3), Léon Bérard Cancer Center , Lyon, France.
Insights
A new whole-blood assay monitors immune cell function in breast cancer patients. It reveals significant immune cell alterations even at early stages, highlighting potential for treatment monitoring.
Area of Science:
- Immunology
- Oncology
- Clinical Diagnostics
Background:
- Monitoring immune cell function is crucial but challenging in routine clinical settings.
- Breast cancer (BC) progression is associated with immune system dysregulation.
Purpose of the Study:
- To develop and validate a whole-blood assay for assessing peripheral innate and adaptive immune cell function.
- To identify immune cell functional alterations in breast cancer patients at various disease stages.
Main Methods:
- Whole-blood assay with short-term stimulation, intracellular cytokine staining, and multi-parametric flow cytometry.
- Analysis of Natural Killer (NK) cells, dendritic cells (DCs), monocytes, and T cell subpopulations.
- Comparison between healthy women and breast cancer patients.
Main Results:
- Significant functional alterations in innate immune cells (DCs, monocytes, NK cells) and T cells observed in BC patients, even at the primary tumor stage.
- Reduced TNFα production by DCs and monocytes, decreased IFNγ by NK cells, and impaired T cell cytokine production (IL-2, IFNγ, IL-21) were noted.
- Immune dysfunctions were more pronounced in advanced disease stages and at first relapse, with increased IL-12p40 production by monocytes.
Conclusions:
- The developed whole-blood assay effectively detects immune cell dysfunctions in breast cancer patients.
- Monitoring both innate and adaptive immunity is vital for evaluating cancer-related immune alterations.
- This assay holds promise for monitoring treatment response, especially in immunotherapeutic strategies.
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.
