Figure 6

CCL5 is involved in the recruitment of Treg cells by c-FOXP3 in vitro and in vivo. (a) FACS analysis. Treg cells recruitment by Foxp3-overexpressed Panc-1 and AsPC-1 cell lines with the CCL5 neutralized antibody for 24-h relative to those with IgG. Treg cells recruitment by Foxp3 knockdown MIA PaCa-2 and BxPC-3 cell lines treated with recombinant CCL5 for 24-h relative to those with IgG. Percentages of recruited Treg cells from peripheral blood mononuclear cells (PBMCs) were shown (mean±s.e.m., n=3; *P<0.05 by Student’s t-test). (b) The cell count of the recruited Treg cells (mean±s.e.m., n=5; *P<0.05 by Student’s t-test). (c) Experimental scheme for subcutaneous carcinoma model of C57BL/6 mice: CCL5 antibody or isotype IgG was injected intratumoral (20 μg per mouse, twice a week). (d, e) FACS analysis of Treg cells recruitment into tumor microenvironment of the four groups was measured. Inhibition rate=(IgG-CCL5Ab) group/IgG group. (f) FACS analysis of CD8⁺T cells apoptosis in tumor microenvironment of the four groups was measured. (g) Tumor growth was evaluated by measuring tumor volumes and growth inhibition rate, and compared by one-way analysis of variance with Bonferroni post-hoc test (**P<0.01). Inhibition rate=(IgG-CCL5Ab) group/IgG group.