Table III.

Simultaneous generation of ORF1- and ORF2-derived epitopes by transfection of NA8-MEL and COS-7 cells with plasmids encoding NY-ESO-1 and LAGE-1 sequences

Cell LinePlasmidaTNF Production by Specific CTLb
Clone LAU 156 NY-ESO-1/5Clone LAU 50 CAMEL/4Line LAU 342 CAMEL
Control+ P 198 864 570
LAGE-1S 101 26 30
LAGE-1L 38 67 19
NY-ESO-1 60 116 21
CAMEL-mini GFP2 680 10
NY-ESO-1-mini GFP 60 34
Control+ P 200 1016 200
LAGE-1S 285 472 40
LAGE-1L 84 752 24
NY-ESO-1 186 800 65
CAMEL-mini2 1184 888
NY-ESO-1-mini 384 84
  • a Cells were transfected with plasmids containing full length cDNAs for NY-ESO-1, LAGE-1S, and LAGE-1L or mini-gene versions of the CAMEL1–11 and NY-ESO-1157–165 epitopes (CAMEL- and NY-ESO-1-mini). Cells transfected with empty vector alone in the absence (control) or presence (control + P) of the relevant antigenic peptide were used as negative and positive controls, respectively. COS-7 cells were cotransfected with an HLA-A2 containing plasmid.

  • b Transfected cells were incubated with clone LAU 156 NY-ESO-1/5 (specific for NY-ESO-1157–165/ORF1), clone LAU 50 CAMEL/4, and line LAU 342 CAMEL (specific for CAMEL1–11/ORF2), and TNF release was measured by a bioassay as described in Materials and Methods. Values greater than 3-fold the corresponding background value were considered positive and are shown underlined.