Targeting myeloid-inflamed tumor with anti-CSF-1R antibody expands CD137+effector T-cells in the murine model of pancreatic cancer

BackgroundThe pancreatic cancer vaccine, GVAX, induces novel lymphoid aggregates in the otherwise immune quiescent pancreatic ductal adenocarcinoma (PDAC). GVAX also upregulates the PD-1/PD-L1 pathway, and a pre-clinical model demonstrated the anti-tumor effects of combination GVAX and anti-PD-1 antibody therapy (GVAX/PD-1). Resistance to GVAX was associated with an immune-suppressive myeloid cell infiltration, which may limit further therapeutic gains of GVAX/PD-1 therapy. The expression of CSF-1R, a receptor important for myeloid cell migration, differentiation and survival, and the effect of its therapeutic blockade in the context of GVAX in PDAC has not been investigated.MethodsLymphoid aggregates appreciated in 24 surgically resected PDAC from patients who received one dose of neoadjuvant GVAX were analyzed with multiplex immunohistochemistry. Flow cytometry analysis of tumor infiltrating T-cells in a murine model of PDAC was performed to investigate the therapeutic effects and mechanism of anti-CSF-1R/anti-PD-1/GVAX combination immunotherapy.ResultsHigh CSF-1R expression in resected PDAC from patients who received neoadjuvant GVAX was associated with a higher myeloid to lymphoid cell ratio (p<0.05), which has been associated with poorer survival. This higher CSF-1R expression was associated with a higher intra-tumoral infiltration of immature dendritic cells (p<0.05), but not mature dendritic cells (p=0.132). In the pre-clinical murine model, administering anti-CSF-1R antibody prior to and after GVAX/PD-1 (pre/post-CSF-1R + PD-1 + GVAX) enhanced the survival rate compared to GVAX/PD-1 dual therapy (p=0.005), but administering anti-CSF-1R only before GVAX/PD-1 did not (p=0.41). The pre/post-CSF-1R+PD-1+GVAX group also had higher intra-tumoral infiltration of PD-1+CD8+ and PD-1+CD4+ T-cells compared to PD-1/GVAX (p<0.001). Furthermore, this regimen increased the intra-tumoral infiltration of PD-1+CD137+CD8+, PD-1+CD137+CD4+ and PD-1+OX40+CD4+ T-cells (p<0.001). These PD-1+CD137+CD8+ T-cells expressed high levels of interferon- (median 80-90%) in response to stimulation with CD3/CD28 activation beads, and this expression was higher than that of PD-1+CD137-CD8+ T-cells (p<0.001).ConclusionsThe conversion of exhausted PD-1+ T-cells to CD137+ activated effector T-cells may contribute to the anti-tumor effects of the anti-CSF-1R/anti-PD-1/GVAX combination therapy. Anti-CSF-1R antibody with anti-PD-1 antibody and GVAX have the potential be an effective therapeutic strategy for treatment of PDAC.