Cell - 8 September 2016

Figure S7.gdT Cells Do Not Alter Myeloid Cell Infiltration or Function in PDA and Localize withabT Cells in the TME, Related toFigure 7
(A) WT and Tcrd–/–mice were orthotopically implanted with KPC-derived pancreatic tumor cells. Tumors were harvested at 3 weeks and analyzed by flow cy-
tometry. CD11b+ myeloid cells were gated and tested for co-expression of Ly6C and Ly6G. Representative contour plots and quantitative data from 5 mice are
shown.
(B and C) CFSE-labeled splenic CD3+T cells were either unstimulated, or stimulated withaCD3/aCD28 alone or in co-culture with orthotopic PDA-infiltrating (B)
MDSC or (C) TAMs (5:1 ratio) derived from WT or Tcrd–/–mice.aPD-L1 was added to select co-culture wells. T cell proliferation was determined by dilution of
CFSE on flow cytometry.
(D and E) CD3+T cells were either unstimulated, or stimulated withaCD3/aCD28 alone or in co-culture with orthotopic PDA-infiltratinggdT cells (d) MDSC or (e)
TAMs (5:1 ratio) +/
aPD-L1 (10mg/ml). T cells activation was determined by expression of TNF-a.
(F and G) Human PDA (F) and orthotopic KPC (G) tumors were co-stained for CD11b/TCRabor TCRgd/TCRab. The closest distance between eachabT cell and
CD11b+myeloid cell orgdT cell, respectively, were calculated. Representative high and low power images and quantitative data are shown. 10 low power fields
were examined per pancreas.
(H and I) Orthotopic KPC tumors were co-stained for (H) DAPI, TCRgd, PD-L1, and TCRabor (I) DAPI, CK19, PD-L1, and TCRaband imaged by confocal mi-
croscopy. Two representative images of each combination is shown.