Functional characterization of transferred TCR transgenic CD8⁺ T cells. P14 CD8⁺ T cells were isolated 5 or 6 days after transfer from the spleens of H8-bcCML mice, C57BL/6 controls, and LCMV-infected mice. (A) The production of IFN-γ, TNF-α, and IL-2 by p14 CD8⁺ T cells was determined in the supernatant with a cytometric bead array assay. Results are given as mean ± SEM of 3 to 10 samples per group, pooled from 4 independent experiments. (B) Splenocytes were analyzed in a ⁵¹Cr-release assay (gp33-pulsed target cells [filled symbols]; nonpulsed target cells [open symbols]). CTL activity is given as mean ± SEM of 4 H8-bcCML mice (●), C57BL/6 controls (♦), and LCMV-infected mice (■). One representative experiment of 2 is shown. (C) FACS analysis of blood of H8-bcCML mice before and 6 days after adoptive transfer of p14 CD8⁺ T cells and control H8-bcCML mice. One representative staining of 5 independent experiments is shown. (D) Isolated p14 CD8⁺ T cells were restimulated in vitro. ³H-thymidine incorporation of isolated p14 CD8⁺ T cells is shown as proliferation index (mean ± SEM of 4-10 mice per group). Results are pooled from 3 independent experiments.