Figure 4.
CD40L promotes M2 to M1 macrophages and IL12 production. Human monocytes were co-cultured with MiaPaCa2 pancreatic cancer tumor cells infected with LOAd700, LOAd(−) or uninfected cells. Cells and supernatants were collected and analyzed by flow cytometry and Meso Scale at day 3 and 5. The experiment was repeated using three different healthy donor-derived monocytes and the figures show pooled data. CD11b+ myeloid cells were gated and the CD163 (M2) macrophages were compared as % positive cells (a) or the MHC-II++ cells were analyzed as fold change from co-cultures with uninfected cells (b, c) granulocyte–macrophage colony-stimulating factor, IL12, IL8 and IL6 levels are shown in the figure. Immunodeficient Nu/Nu mice lacking T lymphocytes but with sustained myeloid cells were implanted with human Panc01 cells. Palpable tumors were injected with mLOAd700, LOAd(−) using 1 × 109 infectious units (IU) per mouse (intratumoral injection; n=3/group). (d) At 48 h post injection, the tumors were measured, and the murine tumor biopsies analyzed by flow cytometry for F4/80+CD11b+CD206− (M1) or F4/80+CD11b+CD206+ (M2) macrophages (e). Error bars represent s.e.m. and statistical significant differences were calculated using Student's t-test.