1. CD1dhiCD5+ B cells are expanded in pancreatic neoplasia and are functionally important for sustaining growth of KrasG12D-PDEC in vivo.
CD1dhiCD5+ B cells are expanded in pancreatic neoplasia and are functionally important for sustaining growth of KrasG12D-PDEC in vivo. A, quantification of flow cytometric analysis of plasma cells from spleens, mesenteric lymph nodes (MLN), and pancreata of p48Cre (control) or KC mice. Cells were analyzed for the presence of markers CD19, B220, and CD138 (n = 5 p48Cre, n = 5 KC). B, quantification of flow cytometric analysis of immune cells from pancreata of p48Cre (control) mice, KC mice (2.5 mo), or KrasG12D-PDEC orthotopic lesions (2 weeks), as indicated. After gating on CD19 and CD1d populations, cells were analyzed for the presence of CD5 marker (n = 8 p48Cre, n = 8 KrasG12D-PDEC, n = 8 KC). C, sections from orthotopic pancreatic grafts 2 weeks after GFP-KrasG12D-PDEC implantation into WT or μMT mice were stained with anti-GFP antibody. Where indicated, μMT mice were reconstituted with WT CD19+CD1dhiCD5+ or with CD19+CD1dloCD5− 2 days prior to orthotopic implantation. Representative images are shown. Scale bars, 100 μm. D, graph depicts quantification of the data from C indicating the average fraction of GFP+ area per field of view (FOV) of the implant (10 fields of view per animal; n = 12 WT, n = 11 μMT, n = 9 μMT+ CD1dloCD5−, n = 9 μMT+ CD1dhiCD5+, animals). E, sections from orthotopic pancreatic grafts 2 weeks after GFP-KrasG12D-PDEC implantation into WT or μMT mice were stained with H&E or anti-GFP antibody. Where indicated, μMT mice were reconstituted with WT B cells or with Il10−/− B cells 2 days prior to orthotopic implantation. Representative images are shown. Scale bars, 100 μm. F, graph depicts quantification of the data from E indicating the average fraction of GFP+ area per field of view of the implant (10 fields of view per animal; n = 14 WT, n = 12 μMT, n = 12 μMT+WT B cell, n = 12 μMT+Il10−/− B cell animals). Error bars indicate SD; P values were determined by the Student t test (unpaired, two-tailed); *, P < 0.05; **, P < 0.01; ***, P < 0.001; NS, not significant.
Yuliya Pylayeva-Gupta et al. Cancer Discov 2016;6:
©2016 by American Association for Cancer Research