Orthotopic PDAC growth is regulated by B cells and FcRγ-positive myeloid cells. Orthotopic PDAC growth is regulated by B cells and FcRγ-positive myeloid.

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Orthotopic PDAC growth is regulated by B cells and FcRγ-positive myeloid cells. Orthotopic PDAC growth is regulated by B cells and FcRγ-positive myeloid cells. A, murine PDAC cell lines, derived from Pdx-Cre; LSL-KrasG12D mice harboring an Ink4a/Arfflox/+ allele (Ink4 2.2) or a p53flox/+ allele (p53 2.1.1), were injected orthotopically into the pancreas, and subsequent tumors evaluated by FACS for CD45-positive cellular infiltrates. Graph shows the percentage of CD45+ cells as a percentage of viable cells. B, representative CD19+MHCII+ FACS plot gated on viable CD45+ cells from tumor-naïve pancreas and Ink4 2.2-implanted PDAC tumors in syngeneic JH+/− and JH−/− mice evaluated 28 days after implantation. C, Ink4 2.2 and p53 2.1.1–derived PDACs were quantitatively evaluated in syngeneic JH+/− and JH−/− mice for tumor area in serial H&E-stained tissue sections reflecting tumors isolated on days 14, 21, and 28 after implantation. On right, representative photomicrographs reflecting immunodetection of αSMA in end-stage Ink4 2.2 and p53 2.1.1–derived orthotopic PDAC tumors. D, Ink4 2.2 and p53 2.1.1–derived PDACs were quantitatively evaluated in syngeneic FcRγ+/− and FcRγ−/− mice for tumor area in serial H&E-stained tissue sections reflecting tumors isolated on days 14, 21, and 28 after implantation. Right, representative photomicrographs reflecting immunodetection of αSMA in end-stage Ink4 2.2 and p53 2.1.1–derived orthotopic PDAC tumors. E, qRT-PCR analysis from cDNA reflecting FACS-purified macrophages (MØ = CD45+CD11b+MHCII+F4/80+Ly6C−) or DC (CD45+CD11b+MHCII+F4/80−CD11c+) from Ink4 2.2 PDAC–derived tumors harvested from day 21 FcRγ+/− (n = 10) or FcRγ−/− (n = 8) mice. Displayed are fold changes in genes that reached statistically significant differences. Data are compiled from two independent experiments. For graphs in A, C–E, statistical means are shown, with P values determined by either Student t test or one-way ANOVA when analyzing more than two groups, with *, P < 0.05; **, P < 0. 0.01; ***, P < 0.001. For graphs in C and D, each data point reflects mean tumor size from one mouse based on quantitative morphometry of 5 FFPE sections, resulting from 3 independent experiments. Andrew J. Gunderson et al. Cancer Discov 2016;6:270-285 ©2016 by American Association for Cancer Research