Figure 4.
Translational investigation of patient T cells treated with glofitamab plus chemotherapy regimens indicates sustained T-cell functionality over prolonged treatment cycles. (A) Clinical trial design for the glofitamab plus GemOx combination (NCT04408638) outlines the treatment schedule and blood sampling time points. (B) Absolute CD3+ T-cell counts in 10 treated patients over time. (C) Experimental design of the ex vivo T-cell assay from patient blood. T cells were isolated as displayed, including glofitamab-mediated tumor cell killing. Glofitamab was used at 1 pM and 1 nM concentration. (D) Glofitamab-mediated tumor cell killing assay using isolated patient CD8+ T cells is assessed. Mean values ± SEM are displayed, with each dot representing an individual patient. (E) Flow cytometry analysis for CD25 and 4-1BB expression on CD8+ T cells after a 24-hour ex vivo killing assay. (F) Cytokine analysis in the supernatant at the 24-hour end point. (G) Design of the clinical trial for the glofitamab plus R-CHOP combination (NCT03467373), indicating the treatment schedule and blood sampling time points. (H) Absolute CD3+, CD8+, and CD4+ T-cell counts in 5 treated patients over time. (I) Glofitamab-mediated tumor cell killing assay using the same experimental setup as in C. (J) Flow cytometry analysis for CD25 and 41BB expression on CD8+ T cells after a 24-hour ex vivo killing assay. (K) Cytokine analysis in the supernatant at the 24-hour end point. C1D1, cycle 1 day 1; E:T, effector to target; IFN-γ, interferon gamma; PBMC, peripheral blood mononuclear cell; Post, after infusion; Pre, before infusion.

Translational investigation of patient T cells treated with glofitamab plus chemotherapy regimens indicates sustained T-cell functionality over prolonged treatment cycles. (A) Clinical trial design for the glofitamab plus GemOx combination (NCT04408638) outlines the treatment schedule and blood sampling time points. (B) Absolute CD3+ T-cell counts in 10 treated patients over time. (C) Experimental design of the ex vivo T-cell assay from patient blood. T cells were isolated as displayed, including glofitamab-mediated tumor cell killing. Glofitamab was used at 1 pM and 1 nM concentration. (D) Glofitamab-mediated tumor cell killing assay using isolated patient CD8+ T cells is assessed. Mean values ± SEM are displayed, with each dot representing an individual patient. (E) Flow cytometry analysis for CD25 and 4-1BB expression on CD8+ T cells after a 24-hour ex vivo killing assay. (F) Cytokine analysis in the supernatant at the 24-hour end point. (G) Design of the clinical trial for the glofitamab plus R-CHOP combination (NCT03467373), indicating the treatment schedule and blood sampling time points. (H) Absolute CD3+, CD8+, and CD4+ T-cell counts in 5 treated patients over time. (I) Glofitamab-mediated tumor cell killing assay using the same experimental setup as in C. (J) Flow cytometry analysis for CD25 and 41BB expression on CD8+ T cells after a 24-hour ex vivo killing assay. (K) Cytokine analysis in the supernatant at the 24-hour end point. C1D1, cycle 1 day 1; E:T, effector to target; IFN-γ, interferon gamma; PBMC, peripheral blood mononuclear cell; Post, after infusion; Pre, before infusion.

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