Skip to main content
. 2021 Apr 5;9(4):e001292. doi: 10.1136/jitc-2020-001292

Figure 7.

Figure 7

oHSV-1 expressing PD-L1 BiTEs activates endogenous ascites T cells to kill ascites tumor cells and macrophages. Total unpurified ascites cells from different patients were infected with parental oHSV-1 or BiTE expressing oHSV-1 at an MOI of 1 for 4 days in normal serum medium or in the presence of autologous fluid (50% v/v). Endogenous T cell activation (CD3+ CD25+) (A) and PD-L1 expression (B) were determined by flow cytometry. Cytotoxicity of CD206+ (C) and CD163+ (D) ascites macrophages in the presence of autologous ascites fluid (50% v/v) were analyzed by staining in the CD11b+/CD64+ population. (E.) Histograms of surface CD25 expression on endogenous ascites T cells (CD3+) and CD206 and CD163 expression on ascites macrophages (CD11b+/CD64+) infected with parental oHSV-1 (yellow), BiTE expressing oHSV-1 or incubated with free PD-L1 BiTEs (40 nM). Untreated: pink, PD-L1 scFv: blue, PD-L1 NB: cyan. Statistical significance was assessed by two-way analysis of variance followed by Bonferroni post hoc analysis. Significance was assessed versus untreated cells within the relevant group (*p<0.05, **p<0.01 and ***p<0.001). BiTE, bispecific T cell engager; NB, nanobody; oHSV-1, oncolytic herpes simplex virus-1; PD-L1, programmed death-ligand 1; scFv, single-chain variable fragment