Functional analysis of retrogenic and transduced peripheral T cells. (A) Retrogenic splenocytes expressing each of the TCRs gated on donor Thy1.2⁺CD8⁺ cells, and TCR-transduced P14 splenocytes gated on transduced (Vβ10⁺) CD8⁺ T cells, were analyzed for expression levels of the retrogenic TCRβ chain, Vβ10, and for tetramer binding. Data are representative of at least 16 mice per TCR. Two samples for each enhanced affinity receptor are included to show the typical range of variation between mice. (B) Retrogenic T cells and TCR-transduced P14 T cells were stimulated with WT1 peptide and expanded for 1 week in vitro and then assayed for functional avidity by measuring interferon γ production in response to Ag-presenting cells pulsed with decreasing concentrations of WT1 peptide. TCR surface expression is shown for each T-cell population at the time the functional assay was performed.