Figure 4. Wild-type (WT) Trp2/K^b-specific cells are capable of an initial response to Trp2 similar to that of Dct^-/- cells.

WT and Dct^-/- mice received intravenous injections of TriVax with 200 μg Trp2 peptide. Tetramer enrichment was used to enumerate Trp2/K^b-specific cells and assess their phenotype at the indicated time points following immunization (A–C, E). The ratio between the mean experimental PE median fluorescence intensity (MFI) of Trp2/K^b-specific cells in WT mice relative to Dct^-/- mice is plotted in D, with each symbol representing one experiment comprising two to five individual mice. Data are compiled from three or more experiments in A, C, and D. Representative flow plots from 1 day four experiment are shown in B, and the same representative day four experiment is shown in E. Squares indicate male animals; the dotted line indicates the average naïve precursor frequency from the spleen and lymph nodes. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 by one-way ANOVA with Sidak’s multiple comparisons test (performed on log-transformed data in [a]).

Figure 4—figure supplement 1. Tetramer staining kinetics and response to peptide immunization.

WT and Dct^-/- mice received intravenous injections of 200 μg Trp2 peptide as part of TriVax (A) or alone (C–E). (A) Histograms of tetramer staining for Trp2/K^b-binding cells at the indicated day after TriVax. (B) Histogram and quantification of tetramer staining for B8R/K^b-binding cells at day 7 after TriVax with B8R peptide are shown as a control. (C–E) Tetramer enrichment was used to enumerate Trp2/K^b-specific cells and assess their phenotype at the indicated time points following peptide injection; males are indicated by square symbols. Data from representative experiments at each time point are shown in (A). Data in C–E are compiled from multiple experiments. ***p<0.001, ****p<0.0001 by one-way ANOVA with Sidak’s multiple comparisons test.