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. Author manuscript; available in PMC: 2016 Nov 1.
Published in final edited form as: Eur J Immunol. 2015 Aug 3;45(11):3140–3149. doi: 10.1002/eji.201545749

Figure 3. The CD27:CD70 interaction influences immunodominance.

Figure 3

A. Schematic of experimental design where WT or CD27−/− OT1 T cells were transferred into mice 18–24 hours before immunization with LPS matured peptide-pulsed WT BMDCs. 5–6 days post immunization mice were sacrifice and stained with peptide specific tetramer to evaluate the ability of endogenous peptide specific T cells to compete with the transferred cells. B. Example flow plots of the frequency of endogenous (CD45.2) tetramer staining cells over the course of 500–5000 transferred WT OT1 T cells (CD45.1). Numbers in lower right quadrant represent the percent tetramer staining T cells out of total endogenous T cells. The staining and gating strategy shown was used in the calculation of endogenous T cell numbers in C–E. C. Total number of endogenous SIINFEKL specific T cells after SIINFEKL loaded BMDC immunization followed by no OT1 T cell transfer (white bar), 500 WT or CD27−/− OT1 T cells transferred (light gray bars), 1000 WT or CD27−/− OT1 T cells transferred (medium gray bars), or 5000 WT or CD27−/− OT1 T cells transferred. Cells were visualized using flow cytometry and gated on B220-, CD8+, CD44+, tetramer+ cells and the congenic marker as described in part B. This experiment was done more than ten times with 3–7 mice per group. D. Total number of endogenous SAINFEKL specific T cells post SAINFEKL loaded BMDC immunization. Done exactly as described in C and repeated at least four times with 3–7 mice per group. E. Total number of endogenous SIYNFEKL specific T cells after SIYNFEKL loaded BMDC immunization followed by OT1 T cell transfer of 0 (white), 5000 (light gray), 20000 (dark gray), or 50000 (black) cells. Experiments in part E were repeated at least twice with 3 mice per group with similar results. All experiments in the figure are representative and one asterisk represents statistically significant differences in the lines as evaluated by two-way ANOVA with a p value <0.05, two asterisks represent a p-value <0.01. Data shown is the mean +/−SEM.

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