The following antibodies were used in this investigation: Treatment grade antibodies Ch14.18/CHO and rituximab were provided by Great Ormond Street Hospital pharmacy. Mouse anti-human CD3-PE/Cy7 (Biolegend 3000316; clone HIT3a), mouse anti-human TCR Vδ1-FITC (Thermo Scientific TCR2730; clone TS8.2), mouse anti-human TCR Vδ2-PE (BioLegend 331408; clone B6), mouse anti-human CD45-APC (Biolegend 304012; clone HI30), mouse anti-human CD45-APC (Biolegend 304006; clone HI30), rat anti-human/mouse CD11b (Biolegend 101228; clone M1/70), mouse anti-human IFNγ-APCCy7 (Biolegend 502530; clone 4S.B3). Live/dead staining was performed using ZombieTM Yellow fixable viability kit (Biolegend 423104), mouse anti-ganglioside GD2-PE (Biolegend 357304, clone 14G2a). Compensation was carried out using single-color controls and eBioScience OneComp eBeads (eBioScience 01-1111). When experiments regarding comparative GD2 expression of neuroblastoma and Ewing's sarcoma cell lines were being carried out, 0.5 × 106 cells were resuspended in 100 μL of buffer and labeled with 3μL of antibody (final concentration 1.5 μg/mL) for 10 min in the fridge before washing twice in PBS. Flow cytometry analysis was carried out on BD LSRII or BD FACSAria flow cytometers and results were analyzed using FlowJo vX.0.7.
Fisher J.P., Flutter B., Wesemann F., Frosch J., Rossig C., Gustafsson K, & Anderson J. (2015). Effective combination treatment of GD2-expressing neuroblastoma and Ewing's sarcoma using anti-GD2 ch14.18/CHO antibody with Vγ9Vδ2+ γδT cells. Oncoimmunology, 5(1), e1025194.
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