Extended Data Fig. 5: SAA regulate the phenotype of tumor-infiltrating T cells.

a) Saa^+/+ or Saa^−/− mice were orthotopically injected with 5e5 PDAC.69 cells. Twenty days later, tumors were harvested and analyzed by CyTOF. tSNE plots from 5000 downsampled CD45⁺ cells were generated and clusters identified using R software. b) Heat map displaying marker expression for each cluster. c) tSNE plots were subsetted on CD3⁺ T cell clusters. d) Expression of immunoregulatory markers on T cell clusters. e) Quantification of CD8⁺Perforin⁺ cells by CyTOF. Data are representative of n = 1 experiment. n = 4 Saa^+/+ and n = 5 Saa^−/− mice. Mann Whitney test performed. f) Quantification of CD3⁺CD8⁺PD1⁺TIM3⁺LAG3⁺ cells by flow cytometry in PDA.69, 7940B, and 6694.c2 subcutaneous tumors at 27-33 days post tumor injection into Saa^+/+ (n = 7 for PDA.69, n = 5 for 7940B, 6694c2) or Saa^−/− (n = 5) mice. n = 1 biological replicate. Mann Whitney tests were performed. g) Experimental schematic for (h-i). C57BL/6 mice (n = 4-6/group) were injected subcutaneously with 1e6 PDA.69 cells on Day 0. T cells were depleted with 0.2mg anti-CD4 and anti-CD8 antibodies on Day −1 and repeated weekly. On Day 28, tumors were formalin fixed, paraffin embedded, and stained for pSTAT1 (yellow), F4/80 (purple), and CK19 (teal). Data shown are from n = 1 biological replicate. h) Quantification of pSTAT1⁺ cells was performed in Visiopharm software. Ordinary one-way ANOVA with Sidak’s multiple comparisons test was performed. i) Representative images of tumors. Scale bars = 100µM. j) Saa^+/+ or Saa^−/− mice were subcutaneously (SC) injected with T cell^low tumors (5e5 PDA.69, 7940B, or 6694.c2. Mean tumor growth curves are shown. Mann Whitney tests were performed. n = 5 mice/group. Data shown are representative of n = 5 biological replicates for PDA.69, n = 1 biological replicate for 7940B and 6694.c2. For e, f, and h, data are presented as mean values +/− SD.

Source data