Mg2b 57

The following antibodies were used: anti-human CD3 (HIT3a, Biolegend), anti-human CD8 (SK1, Biolegend), anti-human NKp30 (P30-15, Biolegend), anti-human HLA-A2 (BB7.2, Biolegend), anti-human HER2 (24D2, Biolegend), anti-mouse TCR-β chain (H57-597, Biolegend), anti-human CD107a (H4A3, Biolegend), anti-human IFN-γ (B27, Biolegend). Corresponding isotype controls mIgG1 (MOPC-21, Biolegend), mIgG2b (MG2b-57, Biolegend) and Armenian Hamster IgG (HTK888, Biolegend) were used. Anti-B7H6 clone 1.18 and corresponding isotype control were from in-house production.^{9 (link)} For CD8⁺ T cell activation, anti-human functional-grade purified anti-CD3 (UCHT1, Biolegend) and anti-CD28 (37.51, Biolegend) were used.

The cytotoxic potential of expanded Vγ9Vδ2 T lymphocytes against DLBCL cells was assayed using propidium iodide (PI, Sangon Biotech Shanghai Co., Ltd.) staining. In brief, CFSE-labeled Vγ9Vδ2 T lymphocytes were co-incubated with DLBCL cells at different effector:target (E:T) ratios of Vγ9Vδ2 T lymphocytes to DLBCL cells (0:1, 5:1, 10:1, and 20:1) at 37°C for 6 h. After cells were washed with PBS, PI solution was added to the co-culture and incubated with the cells for 5 min. Subsequently, the proportion of PI⁺CFSE⁻ cells was quantified using flow cytometry. In some experiments, DLBCL cells were pretreated with z-VRPR-fmk, BPTES or QNZ for 12 h prior to co-culture with Vγ9Vδ2 T lymphocytes. For blocking assays, purified anti-human CD274 antibody (10 μg/ml; 29E.2A3) or isotype control antibody (10 μg/ml; MG2b-57; BioLegend) were included at the time of effector/target co-culture.