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. Author manuscript; available in PMC: 2018 Jan 1.
Published in final edited form as: Clin Cancer Res. 2016 Jun 27;23(1):193–203. doi: 10.1158/1078-0432.CCR-16-0040

Figure 3. Macrophages increase DC function through type I IFN.

Figure 3

(A) A20-bearing WT BALB/c mice (n = 5/group) were administered 100 μg of anti-mCD20 or 100 μg of hIgG on days 11 and 15. Macrophage-depleting reagent (Clophosome, 200 μg/mouse) was administered on day 10 and day 15. (B) Eleven days after Ab treatment, DLN cells were collected, and the IFN-γ ELISPOT assay was performed with medium or irradiated A20 cell restimulation. (C) A20-HA-bearing BALB/c-nude mice (n = 5/group) were treated with anti-mCD20 and/or Clophosome on day 14. Three days later, DCs from DLN were sorted by FACS and then were co-cultured with CL4 T cells. IFN-γ production was measured two days later. (D) Bone marrow-derived dendritic cells or macrophages were co-cultured with A20-HA cells in the presence of anti-mCD20 and/or 2.4G2 for 3 days. IFNβ production was measured by ELISA. ***P<0.0001. (E) A20-bearing WT BALB/c mice (n = 5/group) were administered 100 μg of anti-mCD20 or hIgG on day 12. Three days later, DCs and macrophages from the tumor were sorted by FACS. Total RNA was extracted and used for the real-time PCR detection of IFN-α. (F) A20-HA-bearing BALB/c-nude mice (n = 5/group) were treated with anti-mCD20, or anti-mCD20 and anti-ifnar1, or hIgG on day 14. Three days later, DCs from the DLN were sorted by FACS and then were co-cultured with CL4 T cells. IFN-γ production was measured two days later. (G) A20-bearing WT BALB/c mice (n = 5/group) were administered 100 μg of anti-mCD20 or hIgG on days 12 and 15. Mouse ifnar1-blocking Ab (anti-ifnar1, 100 μg/mouse) was administered twice a week starting on day 11. (H) Eleven days after the Ab treatment, DLN cells were collected, and the IFN-γ ELISPOT assay was performed with medium or A20 cell restimulation. (I) Bone marrow-derived macrophages were co-cultured with Raji cells in the presence of either Rituximab or hIgG for 3 days. Next, the tumor cells were washed out, and CD8+ T cells from Raji-immunized mice were added to the culture. IFN-γ production was measured 3 days later. The mean ± SEM values are shown. Two-tailed Student’s t test was used for statistical analysis.