Extended Data Fig. 9: SAA regulate a tumor-specific T cell response dependent on dendritic cells.

a) Experiment schematic for treating hematopoietic stem cells (HSCs) harvested from Tlr2^+/+ or Tlr2^−/− mice with GM-CSF (0.2µg/mL) and Pam3CSK4 (100ng/mL). b) Quantification of CD11c⁺ DCs by flow cytometry. Significance was determined by ordinary one-way ANOVA with Sidak’s multiple comparisons correction. n = 3 technical replicates per n = 2 biological replicates. c) Study design for (d). HSCs were harvested from Tlr2^+/+ or Tlr2^−/− mice and treated with GM-CSF (0.2µg/mL) for eight days to differentiate them into bone marrow derived dendritic cells (BMDCs). BMDCs were treated with SAA (1µg/mL) for 24 hours. d) IL-6 and IL-10 were detected in supernatant by cytokine bead array. Significance was determined by one-way ANOVA with Sidak’s multiple comparisons correction. Data are representative of n = 3 biological replicates. e) Mice were treated as described in Fig. 3o, and CD8⁺tetramer⁺ cells were quantified by flow cytometry. Significance was determined by Brown-Forsythe and Welch ANOVA test with Dunn’s T3 multiple comparisons test. Data are representative of n = 3 biological replicates. n = 5 mice per group, except for tumor bearing Saa^+/+ mice (n = 3) and tumor bearing Saa^−/− mice (n = 4). f) Representative flow plots of tetramer expression on CD8⁺ T cells. g) Study schematic. Saa^+/+ (n = 7/group) or Saa^−/− mice (n = 8/group) were orthotopically (OT) injected with 5e5 PDA.69 cells on day 0. DC depleting antibody (aDC) was administered on day −2, −1 and twice weekly after tumor implantation. Analysis was performed on Day 20. h) Spleens were analyzed by flow cytometry for DC depletion. n = 6/group for aDC. Mann Whitney tests were performed. i) Tumors were analyzed by flow cytometry for total DC and T cell infiltration. Mann Whitney tests were performed. For b, d, e, h, and i, data are presented as mean values +/- SD.

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