Fig. 6
CCL2 promotes lung tumor growth. A BALB/c mice were intravenously implanted with 1 × 106 67NR cells and 100 ng CCL2 cytokine or PBS vehicle were delivered daily by the intranasal route for 5 days/week for 2 weeks. Two weeks after treatment, mice were sacrificed and the lung weight was determined. Wilcoxon rank sum test, p = 0.006. Mean ± SD. B Photographs of Lungs from CCL2 or PBS treated mice prior to i.v. delivery of 1 × 106 67NR cells. Lungs from mice in Fig. 6a were removed from euthanized mice and representative ones were photographed. PBS-treated mouse lung (a), CCL2-treated mouse lung (b), or tumor-free un-treated lung (c). C Lungs from CCL2-treated mice do not exhibit significant increase infiltrate of CD45+ cells. Mice treated as described in 6A were euthanized; lungs were harvested then prepared for FACS analysis of infiltrating CD45+ leukocytes. Data are reported as % of CD45+ cells total lung cells analyzed. Student’s t-test, p = 0.15. D CCL2 enhanced immune cells infiltrating into lung tumor. BALB/c mice were treated as described in 6a. Two weeks after treatment, the lung tumor microenvironment was analyzed for the infiltration of immune cells by multicolor FACS. Data were analyzed by Student’s t-test. **p < 0.01, n = 5. E CCL2 increases central memory CD8+ T cells but not effector memory T cells. Ea BALB/c mice were treated as described in 6A. Two weeks after treatment, the lung tumor microenvironment was analyzed for memory T cells by multicolor FACS. Data were analyzed by the Student’s t-test. *p < 0.05 vs. PBS controls, n = 5. Eb a representative graph indicating CD44 and CD62L expression on CD8+ T cells from PBS-treated mouse lung or from CCL2-treated murine lung. F CCL2 effects on the polarization of lung macrophages. BALB/c mice were treated as described in 6A. Two weeks after treatment, the lung tumor microenvironment was analyzed by FACS for a) the infiltration of F4/80+ macrophages expressing CD206 (**p < 0.01, n = 5.) and MHC II markers on the cell surface (* p < 0.05, n = 5) (or b) for the intracellular cytokines IFNγ and IL-4 (ns = not significantly different)). Data were analyzed by the Student’s t-test, n = 5