Figure S1 - Defective NKT Cell Activation by CD1d+ TRAMP Prostate Tumor Cells Is Corrected by Interleukin-12 with alpha-Galactosylceramide

PD-L1 blockade does not restore TRAMP-C2 mediated IFNγ secretion in iNKT cells. TRAMP-C2 cells were loaded with α-GalCer vigorously washed and incubated with anti-PDL1 mAbs or isotype controls before freshly isolated liver MNC prestained with mAbs against αβTCr and NK1.1 were added as responder cells. Cocultures were carried out in the presence of Brefeldin A for 6 hours before intracellular IFNγ staining was performed. Data shown are of iNKT cells gated as αβTCR+NK1.1+ cells. Results shown are representative of two experiments. Bottom right histogram: TRAMP-C2 cells do not express PD-L1. TRAMP-C2 cells (bold line) or BM-DC (dashed line) were stained with anti-PD-L1 TRAMP-C2or rat IgG2b isotype control mAbs (dotted line), followed by anti-rat IgG FITC mAbs and analyzed by FACS.
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