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Pleural Effusion of Patients with Malignant Mesothelioma Induces Macrophage- Mediated T Cell Suppression  Lysanne A. Lievense, MD, Robin Cornelissen, MD,

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Presentation on theme: "Pleural Effusion of Patients with Malignant Mesothelioma Induces Macrophage- Mediated T Cell Suppression  Lysanne A. Lievense, MD, Robin Cornelissen, MD,"— Presentation transcript:

1 Pleural Effusion of Patients with Malignant Mesothelioma Induces Macrophage- Mediated T Cell Suppression  Lysanne A. Lievense, MD, Robin Cornelissen, MD, PhD, Koen Bezemer, BSc, Margaretha E.H. Kaijen-Lambers, BSc, Joost P.J.J. Hegmans, PhD, Joachim G.J.V. Aerts, MD, PhD  Journal of Thoracic Oncology  Volume 11, Issue 10, Pages (October 2016) DOI: /j.jtho Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions

2 Figure 1 Expression of signature macrophage phenotype–related genes after culture under standard tumor-inhibitory (M1) or tumor-promoting (M2) conditions or in the presence of pleural effusion (PE) supernatant (n = 6). (A) Gene expression of the general macrophage marker CD68. Expression of the specific M2 markers' scavenger receptor CD163 (B) and interleukin-10 (C). Expression of the activation marker characteristic for the M1 phenotype CD80 (D), expression of the activation marker programmed death ligand 1 (PD-L1) (E), and expression of the proinflammatory marker human leukocyte antigen D–related (HLA-DR) (F). Expression levels are calculated relative to the housekeeping gene β-actin. Journal of Thoracic Oncology  , DOI: ( /j.jtho ) Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions

3 Figure 2 Macrophages suppress T cell proliferation in the presence of pleural effusion (PE). The percentage proliferation of only T cells (control, in malignant pleural mesothelioma cell line supernatant [SN] or in PE) was set at 100% and the T cell proliferation during coculture with macrophages was calculated relative to the basic proliferation of only T cells (A). Coculture with macrophages under control conditions (n = 3) (10% normal human serum) did not show a statistically significant increase or decrease in T cell proliferation (A). Coculture with macrophages in the presence of SN demonstrated an increase in T cell proliferation (n = 6) (*p < 0.05) (A). Coculture with macrophages in the presence of PE induced a suppression of T cell proliferation (n = 6) (p < 0.05, paired Wilcoxon test) (A). (B–F) Representative flow cytometry histograms of the following culture conditions: unstimulated T cell control (B), stimulated T cells (C), stimulated T cells plus macrophages control (D), stimulated T cells plus macrophages SN (E), and stimulated T cells plus macrophages PE (F). MQ, macrophage; FITC-A, fluorescein isothiocyanate; CFSE, carboxyfluorescein succinimidyl ester. Journal of Thoracic Oncology  , DOI: ( /j.jtho ) Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions

4 Figure 3 Concentrations of selected cytokines in pleural effusion (A) supernatants and corresponding malignant pleural mesothelioma (MPM) cell lines supernatants (both n = 6) (B). Cytokine levels were measured using a magnetic bead–based multiplex assay. The prostanoid prostaglandin E2 (PGE2) was measured using an enzyme-linked immunosorbent assay. The depicted concentrations of pleural effusion supernatants and MPM cell line supernatants are undiluted. IL, interleukin; TGF-β, transforming growth factor-β; VEGF, vascular endothelial growth factor; TNF-α, tumor necrosis factor-α; IFN-γ, interferon-γ. Journal of Thoracic Oncology  , DOI: ( /j.jtho ) Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions

5 Figure 4 T cell subsets and tumor-associated macrophages (TAMs) in pleural effusion of 30 patients with malignant pleural mesothelioma. *Tregs are depicted as percentage of CD4 T cells. All other cell populations are depicted as percentages of total live cells (A). Tregs were classified as CD4-positive CD25-positive CD127-negative foxhead box P3–positive cells. TAMs are CD45-positive CD14-positive CD68-positive; M2TAMs are CD206-positive, CD163-positive, or CD206-positive CD163-positive TAMs. (B) Correlation between all TAMs and T cells in the pleural effusion of patients with malignant pleural mesothelioma. T cells are all CD3-positive cells, TAMs are all CD45-positive CD14-positive CD68-positive cells. ***Spearman’s r = –0.90, p < (C) Correlation between all TAMs and Tregs, calculated as a percentage of CD3-positive CD4-positive cells. Tregs were classified as CD4-positive CD25-positive CD127-negative foxhead box P3–positive cells. **Spearman's r = 0.58, p < 0.01. Journal of Thoracic Oncology  , DOI: ( /j.jtho ) Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions

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