Fig. 4.

Lactacystin inhibits cross-presentation of an NY-ESO-1 epitope in LCs/IFN-γ. Cells were preincubated in the presence or absence of lactacystin (40 μM) for 1 h before exposure to NY-ESO-1 IC (3 μg/ml protein and 2 μg/ml specific mAb) or p157-165 (10 μM) and were then tested by ELISPOT for recognition by clone49. As negative controls, IC made with truncated NY-ESO-1 SH protein (AA10-121) or with MAGE-1 protein and their respective specific mAb were used at the same concentration. HLA-A^*0201⁺,NY-ESO-1⁺ melanoma cell line MZ-MEL-19 was used as positive control for clone49. Error bars indicate standard deviation in duplicate wells. Results were representative of at least three independent reproducible experiments.