Defective NKT Cell Activation by CD1d+ TRAMP Prostate Tumor Cells Is Corrected by Interleukin-12 with alpha-Galactosylceramide (original) (raw)
Figure 3
Prostate tumors, primary prostate epithelium, and prostate tumor cell lines express CD1d.
A. Row 1: CD1d expression of TRAMP-C2 cells (bold line) and WT splenocytes (thin line)(dashed line: isotype control), as analyzed by FACS. Top right: CD1d protein expression of TRAMP-C2 cells as determined by Western Blot using rat-anti-mouse CD1d mAb (3C11). Row 2: Fluorescence micrographs show TRAMP-C2 cells stained against mouse CD1d (left) and DAPI (center). B. CD1d expression of primary mouse prostate tumors. Contour plot: Gating of Gr-1−, MHC-classII− cells from CD45− tumor cells. Histogram: CD1d expression on gated tumor cells stained against mouse CD1d (solid) or isotype control (dashed line). C. CD1d expression in human CaP cell lines and primary prostate epithelium stained with human CD1d or isotype controls (dashed lines). D. TRAMP-C2 cells express functional CD1d. TRAMP-C2 were pulsed with α-GalCer or vehicle, washed, loaded with anti-CD1d or isotype control mAbs and incubated with DN32 iNKT hybridoma cells overnight. IL-2 in cell culture supernatants was measured by ELISA. Data shown are representative of 3 experiments (**, P<0.005; ***, p<0.001).