Figure 5.

Targeting CD47 in combination with anti-PD-1 therapy decreases melanoma tumor burden in vivo. (A) Schematic of the melanoma patient cohort therapy regimen. (B, C) scRNA-seq was performed through a 10x genomics system on patient PBMC pre-anti-PD-1 and post-anti-PD-1 therapy (*p<0.05, n=16 (4/group)). (D) Circulating TSP1 was determined through ELISA of human patient plasma pre and post-anti-PD-1 therapy (*p<0.05, n=11–16/group). (E) Patient PBMC stained with antibodies for human CD3, CD8, and CD47 to determine CD47 expression on T cells (F) pre-anti-PD-1 and (G) post-anti-PD-1 therapy through flow cytometry. Data were acquired using BD LSRFortessa X-20 Analyzer and analyzed using FCS Express (*p<0.05, n=24). (H) Schematic of in vivo syngeneic mouse melanoma combination treatment regimen. C57Bl/6 mice were injected subcutaneously with B16 melanoma cells into the outer hind limb. Mice received alternating day intraperitoneal treatments of 10 µM CD47(-) and/or 200 µg anti-PD-1 or their controls over 6 days. (I, J) Tumors were measured three times a week to determine tumor volume (LW²/2). (*p<0.05, n=5). PBMC, peripheral blood mononuclear cells.