Figure 3. Type 2 granulocytes in BM upregulate IL-4 and control myeloid output in response to distal tumor cues.

(a) Proportion of indicated cell type among IL4-eGFP^POS cells in BM of tumor-bearing mice (n=4 mice). Representative of two independent experiments. (b) IL4-eGFP expression in BM eosinophils and basophils from naïve (n=10) and tumor-bearing (n=7) mice. Pooled from two independent experiments. (c) Percentage of lung parenchymal eosinophils and basophils in naïve and tumor-bearing mice, (n=5 mice per group). Representative of two independent experiments. (d) IHC for MCP-8 in indicated tissues. Representative of 12 mice. (e) Tumor burden in KP-bearing mice treated with αFCER1A or isotype antibodies (n=9 mice per group). Pooled from two independent experiments. Scale bar=2mm. (f) Tumor burden in KP-bearing mice treated with αCD200R3 (n=11) or isotype (n=12) antibodies. Pooled from two independent experiments. Scale bar=2mm. (g) Immune populations in tumor-bearing mice treated with αFCER1A or isotype antibodies. n=8 isotype and n=7 αFCER1A-treated mice for mo-macs. n=8 isotype and n=7 αFCER1A-treated mice for monocytes and Ly6c^NEG GMPs. Representative of two independent experiments. (h) Immune populations in tumor-bearing mice treated with αCD200R3 or isotype antibodies. n=11 mice per group for mo-macs. n=12 isotype and 11 αCD200R3-treated mice for monocytes. n=11 isotype and 9 αCD200R3-treated mice for Ly6c^NEG GMPs. Pooled from two independent experiments. (i) IL4-eGFP expression in BM basophils cultured in control or tumor-conditioned media. Representative of two independent experiments. (j) Expression of proteins in lung homogenate of naïve (n=5) and tumor-bearing (n=8) mice. Pooled from two independent experiments. (k) Inflammatory proteins upregulated in plasma of treatment-naïve NSCLC patients (n=29) compared to healthy controls (n=21). Analytes also upregulated in j are indicated in red. One experiment. (l) Expression of IL4-eGFP in BM basophils cultured with indicated cytokines normalized to control media. Two technical replicates per condition, representative of two independent experiments. Unpaired two-tailed Student’s t-test (b,c,g,h,j,k) or Mann-Whitney Test (e,f). Data are mean ± s.d. (a,b,c,g,h,j,k) or median (e,f).