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- W130284078 abstract "Abstract WT1 responsive CTL were generated from CMVpp65 stimulated CD8 cells isolated from three CMV-seropositive HLA-A2 normal donors. CD8 selected T cells were stimulated with dendritic cells pulsed with CMVpp65 overlapping peptides. 10 days after the primary stimulation, CD8 T cells were restimulated with CMV pp65 peptide pulsed PBMC. The CMV-stimulated CTL exhibited CMV-specific but not WT1-specific cytotoxicity after the second stimulation as measured by 51CR release assay (Donor 1=54% and 7%, Donor 2=77% and 14%). However, all CMV-stimulated CTL stained with PE-RMFPNAPYL, a WT1A2 tetramer (Donor 1=2.51%, Donor 2=3.85%, Donor 3=0.49%). EBV-CTL generated from an HLA-A2 donor, and WT1 CTL generated from a non-HLA-A2 donor did not stain with the WT1A2 tetramer (0.02% and 0.01% respectively). Therefore, the WT1A2 tetramer was cross-reactive with pp65-CTL and exhibited HLA-A2 specificity. To enrich the CMV-CTL WT1A2 tetramer staining population, we PE-selected donor 1 PE-WT1 tetramer binding pp65 stimulated CTL. The WT1-selected cells were cloned by limiting dilution. 7 of 8 WT1-selected clones stained with the WT1A2 tetramer ranging from 0.89%-3.56%. WT1-selected clones showed three types of specific cytotoxicity: 1) against pp65-only (3 clones=22-34%), 2) against WT1-only (2 clones=43-44%), or 3) against both WT1 and CMV (1 clone=55% and 24% respectively). 2 of the 8 clones did not exhibit either WT1 or CMV specific cytotoxicity. Only 1 of the 2 clones that exhibit WT1 specific cytotoxicity exhibited WT1-specifc IFN-γ production, as measured by ELISPOT. All clones that displayed CMV-specific cytotoxicity also exhibited CMV-specific IFN-γ production. To our knowledge this is the first description of WT1 cross-reactive, CMV-specific CTL." @default.
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- W130284078 date "2009-04-01" @default.
- W130284078 modified "2023-09-23" @default.
- W130284078 title "Generation of WT1-specific CTL from CMV stimulated CD8 cells (41.32)" @default.
- W130284078 doi "https://doi.org/10.4049/jimmunol.182.supp.41.32" @default.
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