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. Author manuscript; available in PMC: 2017 Apr 1.
Published in final edited form as: J Immunother. 2016 Apr;39(3):105–116. doi: 10.1097/CJI.0000000000000116

Figure 1.

Figure 1

Recognition of WT1 peptide and transfectants by PBL retrovirally transduced to express a WT1 reactive TCR. PBL from 3 HLA-A*0201+ donors were transduced with retroviral particles encoding GFP as a negative control or with the WT1 reactive TCR (TRAV12-1*01; CDR3: CVVNTPPNTDKLIF; TRBV7-2*01; CDR3: CASTPFTSGSGWDEQFF). A. Transduced T cells were cocultured overnight with T2 cells pre-pulsed with titrated concentrations of peptide, and IFNγ in coculture supernatants was measured by ELISA. B and C. COS-7 cells stably expressing high levels of HLA-A*0201 by means of retroviral transduction and antibiotic selection were transiently transfected with GFP cDNA as a negative control or full-length WT1 cDNA. The next day, transduced T cells were cocultured overnight with these transfected cells. IFNγ in coculture supernatants was measured by ELISA (B), and 41BB expression on CD8+ T cells was measured by FACS (C). * indicates >100 pg/ml IFNγ secretion or >5% CD8+ 41BB+ cells by WT1 TCR transduced T cells, >2X background with any negative control target cell, and >2X background of GFP transduced T cells with the same target cell. Bars that reach 2000 pg/ml indicate off-scale IFNγ levels > 2000 pg/ml.