Article Figures & Data
Figures
- FIGURE 1.
Three subsets of DC (CD11c+, CD11c−, and CD11cint) are detected in NSCLC and peritumoral lung tissues using flow cytometry. A, Tumoral and nontumoral lung tissues were analyzed for the presence of CD4+/lineage− (CD3, CD14, CD19, CD35, CD56 and HEA mAbs) cells. B, The different subsets of DC were characterized for their expression of HLA-DR, CD11c, and BDCA-2. C, Total CD4+/lineage− TIDC were enumerated in 22 samples of NSCLC and peritumoral lung tissue and compared with normal peripheral blood. D, The proportion of the three subsets of DC was analyzed in the same samples.
- FIGURE 2.
DC isolated from NSCLC and peritumoral lung tissues have a relatively immature phenotype. A, DC isolated from tumoral and nontumoral lung tissues express low although significantly higher levels of B7-1 and B7-2 molecules compared with blood DC. The expression of CD83 is similarly low in TIDC, in peritumoral DC, and in blood DC. B, CD11cint TIDC correspond to particularly immature DC that express high levels of the immunomodulatory molecule B7-H1. ∗, p < 0.05, significantly different.
- FIGURE 3.
TIDC and peritumoral DC undergo partial phenotypic maturation in response to TLR ligands and have a weak capacity to present Ag to naive T cells. A, PMCs depleted of tumoral cells were activated with LPS, CpG, or R848. Activation of CD11c−, CD11cint, and CD11c+ DC was monitored separately by analyzing the expression of B7-1, B7-2, CD83, and CCR7 by FACS. Data are representative of four independent experiments. B, Supernatants of PMC, either nonactivated or activated with TLR ligands alone or in the presence of blocking IL-10R Ab were harvested, and production of IL-12p40, IFN-α, TNF-α, and IL-10 was quantified by ELISA. ∗, p < 0.05, significant difference. Data are representative of four independent experiments. C, Sorted NSCLC TIDC, either nonactivated or activated for 24 hours with LPS, CpG, and R848 were cocultured with naive T cells (DC to T cell ratio = 1:5) for 4 days. Cultures were pulsed for 24 additional hours with [H3]thymidine and radioactivity incorporation was measured. Data are representative of two independent experiments.
- FIGURE 4.
DC isolated from NSCLC and peritumoral lung tissues migrate in response to chemokines. A, FACS analysis detects significant amounts of CCR-2, CCR-5, CCR-7, and CXCR-3 at the surface of total TIDC. B, When TIDC were incubated in Transwell inserts for 2 h at 37°C, both CD11c+ tumor-infiltrating mDC and CD11c− tumor-infiltrating pDC migrated vigorously toward CCL19 and CXCL12. Migration index was determined by the ratio of specific migration to spontaneous migration in medium alone. Data represent three independent experiments.
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