Figure 5.

A greater fraction of naïve TCR-V cells undergoes proliferation in response to Q489A or G490A. (A–D) Total CFSE labeled splenocytes from TCR-V mice and Thy1.1 congenic mice were mixed together (Thy1.1-spike) and aliquots containing 1×10⁶ TCR-V T cells were transferred into B6 mice. The following day, mice were immunized with the indicated cell lines (A–C) or with the indicated peptide or recombinant vaccinia virus expressing ER-targeted minigene products (D). Three days following immunization, splenocytes were stained with Tet-V, anti-CD8 and anti-Thy1.1. The ratio of CFSE-Hi “UnDivided” TCR-V / CFSE-Hi CD8+Thy1.1+ cells was calculated as in the example shown in (A). (B) Results show the fraction of undivided TCR-V cells in each immunization group. The control group consists of unimmunized mice or mice immunized with Null cells. (C) The percent of naive T cells that divided was calculated by the formula 100*(1-(I/U)) where I = the mean ratio from a group of immunized mice and U = the mean ratio from the group of unimmunized mice. (B–C) Results from two independent experiments were normalized and combined. P values were determined by unpaired t-tests. (D) Data are presented as in B and the percentages shown as in C. For mice immunized with peptide, U = the mean ratio from peptide-I immunized mice. For mice immunized with rVV-ES-V, U = the mean ratio from rVV-ES-I immunized mice. The data are representative of two independent experiments with groups of three mice.