Loss of virus-specific CD8 T-cell reactivity in dispersed TMCs. (A) TMCs were prepared from uninfected fresh human tonsils and were either kept on ice or dispersed in suspension (1 × 106 cells/ml) for 2 hours at 37°C (5% CO2) to simulate tissue exit. IFN-γ responses to titrated amounts of the HLA-class I restricted PepMix CEF standard were analyzed by an IFN-γ ELISPOT assay, and the frequency of CD8 memory T cells in TMCs was determined by flow cytometry. Unpaired Student t test: *P < .05. (B) Virus-specific IFN-γ responses to 0.04 or 0.4 μg/ml of the peptide pool were compared between CD8 memory T cells from TMCs with those of fresh and HD precultured PBMCs of the same individual as presented in (A). Similar results were obtained after repetition of the experiment. Two-way analysis of variance, ***P < .0005. Data represent mean ± SD for triplicate samples. n.s., not significant.
