Fig. 2: CD206hi IMs organize lymphocyte recruitment and antitumor immunity.
From: Chemokine-defined macrophage niches establish spatial organization of tumor immunity
a, Representative flow cytometry plots showing the gating strategy for the identification of CD11b⁺CD64⁺CD206hi/loFOLR2+/− IMs (left) and time course analysis of the depletion of CD206hiFOLR2+ IMs at days 0, 7 and 15 (middle) and a scatter-plot showing the frequency of CD206hiFOLR2+ IM at day 0–15 (right) after intravenous injection of 700 ng DT in Pf4CreCx3cr1DTR mice. Two independent experiments, n = 2 mice per time point. b, Representative images of tumor-burdened lungs (left) and scatter-plot showing the number of surface metastases per lung (right) in Cx3cr1DTR (n = 5) and Pf4CreCx3cr1DTR (n = 5) mice at day 16 after intravenous injection of B16F10 melanoma cells. Shown is one of four independent experiments. Student’s two-tailed t-test with P < 0.0001. mets, metastases. c, Representative H&E-stained lung sections (left) and quantification of number of tumors per lung section (right) in AgerCreERT2KP mice reconstituted with Cx3cr1DTR (n = 5) and Pf4CreCx3cr1DTR (n = 5) BM. Scale bar, 1,000 µm. One of two independent experiments is shown. Student’s two-tailed t-test with P = 0.0008. d, Representative H&E-stained lung sections (left) and quantification of the number of tumors per section (right) in Cx3cr1DTR (n = 5) and Pf4CreCx3cr1DTR (n = 5) mice at day 16 post-intravenous injection of KPAR1.3 adenocarcinoma cells. Scale bar, 1,000 µm. One of four independent experiments is shown. Student’s two-tailed t-test with P < 0.0001. e, Representative immunohistochemistry images of B220 and CD3e antibodies staining (left) and quantification of B cell and T cell numbers (right) in the TME of Cx3cr1DTR (n = 5) and Pf4CreCx3cr1DTR (n = 5) mice at day 16 after intravenous injection with B16F10 melanoma cells. Scale bar, 100 µm. One of two independent experiments is shown. Student’s two-tailed t-test with P = 0.0027 (B cells) or P = 0.0006 (T cells). f, Representative immunohistochemistry images of B220 and CD3e antibodies staining (left) and quantification of B cell and T cell numbers (right) in the lungs of AgerCreERT2KP mice reconstituted with Cx3cr1DTR (n = 5) and Pf4CreCx3cr1DTR (n = 5) BM. Scale bar, 100 µm. One of two independent experiments is shown. Student’s two-tailed t-test with P = 0.0013 (B cells), P = 0.0008 (T cells). g, Representative immunohistochemistry images of B220 and CD3e antibodies staining (left) and quantification of histopathological scores (right) in the lungs of Cx3cr1DTR (n = 5) and Pf4creCx3cr1DTR (n = 5) mice at day 16 after intravenous injection of KPAR1.3 adenocarcinoma cells. Whole-lung images (stitched, scale bar 1,000 µm, top left) and magnified (×4 and ×10) views (scale bar, 100 µm, bottom left). One of three independent experiments is shown. Student’s two-tailed t-test with P = 0.0005. h, ELISA measurements of CXCL9, CXCL10 and CXCL13 in homogenized lungs from Cx3cr1DTR (n = 5) and Pf4CreCx3cr1DTR (n = 5) mice on day 16 after intravenous injection of KPAR1.3 adenocarcinoma. Two independent experiments. Student’s two-tailed t-test with P < 0.0001, P < 0.0001 and P = 0.0398. All data are represented as mean ± s.e.m. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.
