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Non-small Cell Lung Cancer Induces an Immunosuppressive Phenotype of Dendritic Cells in Tumor Microenvironment by Upregulating B7-H3  Thomas Schneider,

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Presentation on theme: "Non-small Cell Lung Cancer Induces an Immunosuppressive Phenotype of Dendritic Cells in Tumor Microenvironment by Upregulating B7-H3  Thomas Schneider,"— Presentation transcript:

1 Non-small Cell Lung Cancer Induces an Immunosuppressive Phenotype of Dendritic Cells in Tumor Microenvironment by Upregulating B7-H3  Thomas Schneider, MD, Hans Hoffmann, MD, PhD, Hendrik Dienemann, MD, PhD, Philipp A. Schnabel, MD, PhD, Alexander H. Enk, MD, Sabine Ring, PhD, Karsten Mahnke, PhD  Journal of Thoracic Oncology  Volume 6, Issue 7, Pages (July 2011) DOI: /JTO.0b013e31821c421d Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

2 FIGURE 1 Expression of B7 molecules by dendritic cells (DC) in tumor and lung tissue. Single cell suspensions were prepared from lung cancer samples (tumor) and from unaffected lung tissue (control). Suspensions were stained with antibodies as indicated and analyzed by FACS. A, To gate for CD11c+ DC, autofluorescent cells were determined first by measuring the fluorescence of the cells in the APC channel. These cells were excluded. Next, the SSChigh/CD11c+ cells were gated and analyzed for typical DC (HLA-DR), granulocyte (CD16), and monocyte (CD14) markers. B, Cells gated as in (A) were further analyzed for expression of B7 markers. The vertical lines depict the border between negative and positive cells as determined by isotype controls and the number in the upper right corner displays the percentage of positive cells among the CD11c+ population. A typical example for one representative patient out of four is shown. Journal of Thoracic Oncology 2011 6, DOI: ( /JTO.0b013e31821c421d) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

3 FIGURE 2 B7-H3 is upregulated in tumor-derived dendritic cells (DC). Upregulation of B7-H3 was found in all lung cancer samples compared with unaffected lung tissue. The figure shows the data of four experiments as individual dots. The mean values are indicated as bars. *Significant difference (t test; *p < 0.05). Journal of Thoracic Oncology 2011 6, DOI: ( /JTO.0b013e31821c421d) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

4 FIGURE 3 Expression of B7 molecules by dendritic cells (DC) in lymph nodes. Single cell suspensions from tumor-infiltrated lymph nodes from lung cancer patients (TuLN) and tumor-free lymph nodes (CoLN) were prepared and stained with the antibodies indicated. The isotype control is shown in the upper left panel and the big square gate indicates the CD11c+ DC population. The upper gate depicts DC positive for the respective B7 markers. The numbers indicate the percentage of marker-positive cells among the CD11c+ population. A typical example for one representative patient is shown. Journal of Thoracic Oncology 2011 6, DOI: ( /JTO.0b013e31821c421d) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

5 FIGURE 4 Tumor-derived dendritic cells (DC) are inferior in stimulating T cells. CD11c+ cells were enriched by magnetobeads from organs as indicated. The upper panel shows a typical FACS plot for DC-specific markers. The lower panel shows cocultures of CD11c+ cells with allogeneic CD4+ T cells. After 96 hours, T cell proliferation was determined by pulsing with 3H-Thymidine. As positive control, CD4+ T cells were stimulated with a mixture of anti-CD3/anti-CD28 antibodies. The figure shows CPM values of three individual experiments as dots and the means are depicted in bars. *Significant difference (t test; *p < 0.05) between samples as indicated. Journal of Thoracic Oncology 2011 6, DOI: ( /JTO.0b013e31821c421d) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

6 FIGURE 5 B7–H3 blockade results in enhanced stimulation of T cells. Dendritic cell (DC)-T cell cocultures were set up as in Figure 4 and anti-B7-H3 or respective isotype control antibodies (αISO) were added. After 96 hours, T cell proliferation was determined by pulsing the cultures with 3H-Thymidine. The figure shows the individual data as dots and the means as bars. *Significant difference (t test, *p < 0.05) between tumor-derived DC and B7-H3-blocked tumor-derived DC. Journal of Thoracic Oncology 2011 6, DOI: ( /JTO.0b013e31821c421d) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

7 FIGURE 6 Tumor-derived dendritic cells (DC) produce less IL-12 but more IL-10. DC were purified with paramagnetic beads and cultivated overnight in medium containing 10 ng/ml LPS. Thereafter, tissue culture supernatant was harvested and analyzed for presence of IL-10 and IL-12p70 as indicated. Shown are the mean values ± SD of three patients. *Significant difference (*p < 0.05) between tumor and lung. Journal of Thoracic Oncology 2011 6, DOI: ( /JTO.0b013e31821c421d) Copyright © 2011 International Association for the Study of Lung Cancer Terms and Conditions

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