Figure 7
Figure 7. Inhibition of lymphoma dissemination by anti-CD47 antibody requires macrophages and blockade of SIRPα. (A) Subcutaneously engrafted DLBCL mice with palpable flank tumors (3 weeks after transplantation) were treated with control IgG or anti-CD47 antibody for 11 days in the presence of liposomal clodronate or liposomal control. Tumor volume was assessed and indicated that clodronate administration partially abrogated anti-CD47 antibody-mediated reduction in tumor volume compared with control (P = .25). No difference was observed between clodronate and liposomal control administration in control IgG-treated mice (P = .62). In mice treated with liposomal control, anti-CD47 antibody partially reduced tumor volume compared with control IgG treatment, but this difference was not statistically significant (P = .49). Statistical analysis was conducted with a 2-way ANOVA test. (B-C) The presence of lymphoma in the bone marrow (B) and peripheral blood (C) of anti-CD47 antibody-treated mice was assessed in respective treatment cohorts. Lymphoma dissemination to the bone marrow and peripheral blood was observed in anti-CD47 antibody-treated mice receiving clodronate macrophage depletion, but not in controls. Statistical analysis was conducted with Fisher exact test. Each treatment group consisted of 3 or 4 mice. (D) Luciferase-labeled Raji cells were coated ex vivo with the indicated primary antibodies, labeled with a fluorescently conjugated secondary antibody, and analyzed for binding by flow cytometry. (E) Raji cells incubated ex vivo with the indicated antibodies were transplanted intravenously into adult NSG mice. One week later, mice were analyzed by bioluminescent imaging for engraftment, and representative mice are shown. (F) The luciferase signal was determined for a cohort of mice transplanted with ex vivo antibody-coated cells where each point represents a single mouse. Blocking anti-CD47 (B6H12.2) antibody inhibited tumor engraftment compared with the nonblocking anti-CD47 (2D3) antibody and antibody controls. Statistical analysis was conducted with Fisher exact test. Data are mean ± SD. *P < .05. **P < .005. ***P < .0005.

Inhibition of lymphoma dissemination by anti-CD47 antibody requires macrophages and blockade of SIRPα. (A) Subcutaneously engrafted DLBCL mice with palpable flank tumors (3 weeks after transplantation) were treated with control IgG or anti-CD47 antibody for 11 days in the presence of liposomal clodronate or liposomal control. Tumor volume was assessed and indicated that clodronate administration partially abrogated anti-CD47 antibody-mediated reduction in tumor volume compared with control (P = .25). No difference was observed between clodronate and liposomal control administration in control IgG-treated mice (P = .62). In mice treated with liposomal control, anti-CD47 antibody partially reduced tumor volume compared with control IgG treatment, but this difference was not statistically significant (P = .49). Statistical analysis was conducted with a 2-way ANOVA test. (B-C) The presence of lymphoma in the bone marrow (B) and peripheral blood (C) of anti-CD47 antibody-treated mice was assessed in respective treatment cohorts. Lymphoma dissemination to the bone marrow and peripheral blood was observed in anti-CD47 antibody-treated mice receiving clodronate macrophage depletion, but not in controls. Statistical analysis was conducted with Fisher exact test. Each treatment group consisted of 3 or 4 mice. (D) Luciferase-labeled Raji cells were coated ex vivo with the indicated primary antibodies, labeled with a fluorescently conjugated secondary antibody, and analyzed for binding by flow cytometry. (E) Raji cells incubated ex vivo with the indicated antibodies were transplanted intravenously into adult NSG mice. One week later, mice were analyzed by bioluminescent imaging for engraftment, and representative mice are shown. (F) The luciferase signal was determined for a cohort of mice transplanted with ex vivo antibody-coated cells where each point represents a single mouse. Blocking anti-CD47 (B6H12.2) antibody inhibited tumor engraftment compared with the nonblocking anti-CD47 (2D3) antibody and antibody controls. Statistical analysis was conducted with Fisher exact test. Data are mean ± SD. *P < .05. **P < .005. ***P < .0005.

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