T cell killing assay
T2 cells engineered with luciferase reporter (Luc) were maintained in
IMDM supplemented with 2mM L-glu, 20% FBS, 2 ug/mL Blasticidin, and
0.25 ug/mL Puromycin. T2 Luc cells were pulsed with MAGE-B2 peptide at
10 µM for 4 hours. Previously frozen TCR-T cells were thawed and mixed
with MAGE-B2 peptide-pulsed T2 Luc cells at various E:T ratio in CTS
OpTmizer T Cell Expansion SFM (Gibco) with 2.5% CTS Immune Cell SR
(Gibco). Cell mixtures were incubated at 5% CO2 and
37°C for 48 hours. Steady-Glo® Luciferase (Promega) was added to cell
mixture according to manufacturer’s protocol. Luminescence was read
using EnVision plate reader (PerkimElmer) 5 minutes after reaction.