Figure 2.gdT Cells Are Highly Prevalent and Exhibit a Uniquely Activated Phenotype in Murine Invasive PDA
(A) C57BL/6-Trdctm1Malmice whosegdT cells express GFP were orthotopically implanted with KPC-derived tumor and imaged by intra-vital two-photon laser-
scanning microscopy at 21 days.
(B) WT mice were orthotopically implanted with KPC-derived tumor cells. On day 21, single-cell suspensions of digested PDA tumors and splenocytes were co-
stained for CD45, CD3, TCRg/d, CD4, and CD8 and analyzed by flow cytometry. Representative contour plots and quantitative data are shown.
(C) WT mice were orthotopically implanted with KPC-derived tumor cells. On day 21 spleen (blue histograms) and PDA-infiltrating (red histograms)gdT cells were
gated and tested for co-expression of select surface activation markers and Vgchains. Representative histogram overlays and summary data from five mice are
shown.
(DāH) Spleen and PDA-infiltratinggdT cells from the same mice were tested for expression of (D) IL-10, (E) IL-17, (F) NKG2D, (G) TLR4, TLR7, and TLR9, and (H)
CCR2, CCR5, and CCR6. Each experiment was repeated at least three times using three to five mice per data point (*p < 0.05, **p < 0.01).
1488 Cell 166 , 1485ā1499, September 8, 2016