Effect of First-Line Treatment on Myeloid-Derived Suppressor Cells’ Subpopulations in the Peripheral Blood of Patients with Non–Small Cell Lung Cancer

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Effect of First-Line Treatment on Myeloid-Derived Suppressor Cells’ Subpopulations in the Peripheral Blood of Patients with Non–Small Cell Lung Cancer Filippos Koinis, MD, Eleni Kyriaki Vetsika, PhD, Despoina Aggouraki, MSc, Eleftheria Skalidaki, MSc, Anna Koutoulaki, PhD, Marianthi Gkioulmpasani, MSc, Vassilis Georgoulias, MD, PhD, Athanasios Kotsakis, MD, PhD Journal of Thoracic Oncology Volume 11, Issue 8, Pages 1263-1272 (August 2016) DOI: 10.1016/j.jtho.2016.04.026 Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions

Figure 1 Mean change in the percentages of the granulocytic myeloid-derived suppressor cell (G-MDSC) subpopulation. Comparison of the mean change in the percentages of G-MDSCs in the peripheral blood of the patients after three cycles of therapy. The decrease observed in the bevacizumab group is significant when compared with the increase documented in the non-bevacizumab group of patients. The bars denote mean values plus or minus the standard error of the mean, and the p values are as determined by the Kolmogorov-Smirnov nonparametric test. For more details see Supplementary Table 1. Journal of Thoracic Oncology 2016 11, 1263-1272DOI: (10.1016/j.jtho.2016.04.026) Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions

Figure 2 Vascular endothelial growth factor receptor (VEGFR) expression in myeloid-derived suppressor cell (MDSC) subpopulations. Representative histogram of flow cytometry analysis of VEGFR expression on different MDSC subpopulations (A). The gates for the histogram are presented on the top of each box. The positive expression of VEGFR (right histogram) is compared with cells without antibody staining (left histogram). Colors in histograms represent the different subpopulations: purple is used for the population of CD15-positive monocytic MDSCs (M-MDSCs), light blue is used for CD15-negative M-MDSCs, and gray is used for granulocytic MDSCs (G-MDSCs). (B) Comparison of the percentages between MDSC subpopulations expressing the VEGFR receptor in the whole blood of the patients. Percentages indicated in the plots represent the percentages of VEGFR expression in the parental population, which are presented inside the brackets. The bars denote mean values for standard error of the mean, which are as determined by the Wilcoxon matched-pairs signed rank test. Journal of Thoracic Oncology 2016 11, 1263-1272DOI: (10.1016/j.jtho.2016.04.026) Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions

Figure 3 Inhibitory role of myeloid-derived suppressor cell (MDSC) subpopulations. Decreased interferon gamma (IFN-γ) levels in the supernatant from cocultures of activated CD3-positive T cells with MDSCs isolated from patients with non–small cell lung cancer detected using enzyme-linked immunosorbent assay. The experiments were performed in duplicate. The data are presented as mean values plus or minus the standard error of the mean from five independent experiments and the p values are as determined by the Friedman test and Mann-Whitney U test. ∗∗p<0.01 and ∗∗∗p<0.001. HLA-DR, human leukocyte antigen–antigen D–related. Journal of Thoracic Oncology 2016 11, 1263-1272DOI: (10.1016/j.jtho.2016.04.026) Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions

Figure 4 Dynamic changes in myeloid-derived suppressor cell (MDSC) subpopulations after three cycles of chemotherapy and their correlation with tumor response. All MDSC subpopulations (A, B, and C) are significantly elevated in patients experiencing progressive disease (PD) after three cycles of chemotherapy. The bars denote mean values. p Values are as determined by the Kolmogorov-Smirnov nonparametric test. Patients experiencing PD after three cycles of treatment had a statistically significant increase in their percentage of CD15-positive M-MDSCs (A). p Values are as determined by the Wilcoxon matched-pairs signed rank test between different time points. HLA-DR, human leukocyte antigen–antigen D–related. Journal of Thoracic Oncology 2016 11, 1263-1272DOI: (10.1016/j.jtho.2016.04.026) Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions

Figure 5 Baseline corrected percentage of circulating myeloid-derived suppressor cells (MDSCs) in each patient after three cycles of treatment. Waterfall plots illustrating the change in the percentage of the three MDSC subpopulations: CD15-positive cells monocytic MDSCs (A), CD15-negative monocytic MDSCs (B), and granulocytic MDSCs (C) in the peripheral blood of each patient. Patients experiencing a partial response (PR), stable disease (SD), or progressive disease (PD) are presented with green, blue, and red tabs, respectively. HLA-DR, human leukocyte antigen–antigen D–related. Journal of Thoracic Oncology 2016 11, 1263-1272DOI: (10.1016/j.jtho.2016.04.026) Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions

Supplementary Figure 1 Journal of Thoracic Oncology 2016 11, 1263-1272DOI: (10.1016/j.jtho.2016.04.026) Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions

Supplementary Figure 2 Journal of Thoracic Oncology 2016 11, 1263-1272DOI: (10.1016/j.jtho.2016.04.026) Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions

Supplementary Figure 3 Journal of Thoracic Oncology 2016 11, 1263-1272DOI: (10.1016/j.jtho.2016.04.026) Copyright © 2016 International Association for the Study of Lung Cancer Terms and Conditions