Antiviral responses are preserved following CD25/71 allodepletion. (A) CMV-specific CD8⁺ T cells are preserved after CD25/71 allodepletion. The figure shows a representative FACS analysis from 1 of 4 D-R pairs demonstrating staining of either unmanipulated PBMCs (left) or CD25/71 allodepleted cells (right) in a HLA-A2–positive, CMV-seropositive donor with a HLA-A2–CMV pp65 pentamer (top right quadrants). The percentages of pentamer-positive cells as a proportion of CD8⁺ cells with isotype subtracted are shown. (B) EBV-specific CD8⁺ T cells are retained after CD25/71 allodepletion. Representative FACS analysis from 1 of 4 D-R pairs demonstrating staining of either unmanipulated PBMCs (left) or CD25/71 allodepleted cells (right) in a HLA-A2–positive, EBV-seropositive donor with a HLA-A2–CLG pentamer (top right quadrants). The percentages of pentamer-positive cells as a proportion of CD8⁺ cells with isotype subtracted are shown. (C) Functional T-cell responses to CMV and EBV are preserved after CD25/71 allodepletion (n = 5). The frequency of cells secreting IFN-γ in response to stimulation with irradiated autologous PBMCs pulsed with a peptide mix from CMV pp65 or autologous EBV LCL was determined by ELISPOT assays. Unmanipulated PBMCs or CD25/71 allodepleted T cells from the same seropositive donors were compared. Line = median, box = 25th-75th centile, error bars = minimum, maximum. (D) T cells responses to adenovirus are preserved after CD25/71 allodepletion (n = 4). The frequency of cells secreting IFN-γ in response to stimulation with irradiated autologous PBMCs transduced with an adenoviral vector (Ad5f35-GFP) was determined by ELISPOT assay. Unmanipulated or CD25/71 allodepleted T cells from the same seropositive donors were compared.