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Pleural Effusions from Patients with Mesothelioma Induce Recruitment of Monocytes and Their Differentiation into M2 Macrophages  Anne-Laure Chéné, MD,

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Presentation on theme: "Pleural Effusions from Patients with Mesothelioma Induce Recruitment of Monocytes and Their Differentiation into M2 Macrophages  Anne-Laure Chéné, MD,"— Presentation transcript:

1 Pleural Effusions from Patients with Mesothelioma Induce Recruitment of Monocytes and Their Differentiation into M2 Macrophages  Anne-Laure Chéné, MD, Sènan d’Almeida, PhD, Thibaut Blondy, MS, Julie Tabiasco, PhD, Sophie Deshayes, Jean-François Fonteneau, PhD, Laurent Cellerin, MD, Yves Delneste, PhD, Marc Grégoire, PhD, Christophe Blanquart, 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 Effect of C-C motif chemokine ligand 2 (CCL2) present in mesothelioma microenvironment on monocyte migration. Migration of monocytes through transwell filters was evaluated in the presence of pleural effusions (PEs) from patients with mesothelioma (A) or mesothelioma cell culture supernatants (SN) (B) containing various CCL2 concentrations. Using a blocking antibody directed against CCL2 (10 μg/mL), we determined the implication of this chemokine in monocyte migration. (C) Validation of the blocking antibody in standard conditions (2% human serum albumin [HSA]; 10 ng/mL of CCL2 [n = 5]). The effect of the antibody was studied on monocyte migration induced by PEs from patients with MPM (n = 9) (D) or by mesothelioma cell culture SNs (n = 8) (E). r = Spearman’s r; ***p < Journal of Thoracic Oncology  , DOI: ( /j.jtho ) Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions

3 Figure 2 Effect of pleural effusions (PEs) from patients with malignant pleural mesothelioma (MPM) and MPM cell culture supernatants (SNs) on macrophage phenotype. (A) Monocytes were incubated with granulocyte-macrophage colony-stimulating factor (GM-CSF) (20 ng/mL), macrophage colony-stimulating factor (M-CSF) (50 ng/mL), or PEs from patients with MPM (n = 8). Cells were labeled with an anti–CD14–fluorescein isothiocyanate and an anti–CD163-allophycocyanin and analyzed by flow cytometry. (B) Macrophages obtained in (A) were stimulated with lipopolysaccharide (200 ng/mL). Then, interleukin-12 (IL-12) and interleukin-10 levels were measured in SNs using enzyme-linked immunosorbent assay. (C) M-CSF levels in benign PE (BPE) (n = 11) and in PEs from patients with MPM (n = 22). ∗p < (D) Monocytes were incubated for 3 days with M-CSF (50 ng/mL) or PE (n = 8) in the presence or absence of 1 μM GW2580. ∗∗p < (E) Monocytes were incubated with GM-CSF (20 ng/mL), M-CSF (50 ng/mL), or MPM cell culture SNs (n = 8). Cells were labeled with an anti–CD14–fluorescein isothiocyanate and an anti–CD163- allophycocyanin and analyzed by flow cytometry. (F) M-CSF levels in MPM cell culture SNs (n = 14). RFMI, relative mean fluorescence intensity. Journal of Thoracic Oncology  , DOI: ( /j.jtho ) Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions

4 Figure 3 Presence of M2 macrophages in human malignant mesothelioma tumors. Representative results of immunohistological labeling of CD163 in two malignant pleural mesotheliomas (MPMs) and two peritoneal mesotheliomas on a tissue microarray. Arrows indicate CD163-labeled macrophages. Journal of Thoracic Oncology  , DOI: ( /j.jtho ) Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions

5 Figure 4 Effect of M2 macrophages on malignant pleural mesothelioma (MPM) cell proliferation and chemotherapy sensitivity. Monocytes were incubated with macrophage colony-stimulating factor (50 ng/mL) to obtain M2 macrophages. MPM cells were cocultured with increasing amounts of M2 macrophages (10 or 100 M2 macrophages for 5 × 103 MPM cells). (A) MPM cell growth was evaluated using Uptiblue cell counting reagent. Results are expressed as the percentage of MPM cell growth measured in the absence of M2 macrophages (100%). (B) Sensitivity of MPM cells to cisplatin (Cis) (0.8 mg/L) and/or pemetrexed (80 μM). Cell growth was measured after 72 hours of treatment using Uptiblue cell counting reagent. Results are expressed as the percentage of MPM cell growth measured in the absence of treatment (100%). Results are the means plus standard error of the mean of experiments performed on three MPM cell lines. *p < 0.05, ***p < Journal of Thoracic Oncology  , DOI: ( /j.jtho ) Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions

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