Figure 7.

Anti-PD-L1 increases IFN-γ producing CD4 and CD8 T cells in fungal sepsis. Mice had CLP followed three days later by Candida injection.The protocol for administration of anti-PD-1 and anti-PD-L1 as well as the details of splenocyte preparation and incubation were as described in Figure 6.Following overnight stimulation with anti-CD3/CD28, cells were treated with brefeldin for four hours.Cells were then fixed, permeabilized and stained with anti-IFN-γ.Flow cytometry was performed to quantitate the number of IFN-γ⁺ CD4 and CD8 T cells. The number of IFN-γ⁺ CD4 T cells was decreased in two-hit fungal sepsis compared to sham mice and sham mice treated with anti-PD-L1, (P <0.05).Septic mice treated with anti-PD-L1 had an increase in the number of IFN-γ⁺ CD4 and CD8 T cells compared to septic mice treated with saline diluent (P <0.05).CD, cluster of differentiation; CLP, cecal ligation and puncture; IFN-γ, interferon gamma; PD-1, programmed cell death 1; PD-L1, programmed cell death ligand 1.