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. 2016 Dec 12;213(13):3057–3073. doi: 10.1084/jem.20160938

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© 2016 Takamura et al.

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Figure 7. — Prime-pull vaccination with a cognate Ag converted T_EM cells to T_RM cells in the lung. (A–H) Mice were infected i.n. (A) or i.p. (A–E, G, and H) with x31. Some i.p. infected mice were then injected i.n. with PBS, CpG ODN, or CpG ODN + NP_366–374 peptide at the indicated time points. (A) Numbers of NP-specific CD8⁺ T cells in each tissue at day 30 PI. (B) Ratios of CD69⁺, CD49a⁺, and CD103⁺ cells among NP-specific CD8⁺ T cells in the LI/LP and airways. (C) Numbers of NP-specific CD8⁺ T cells in each tissue at day 11 and 30 PI (day 3 and 22 after injection). (D) Expression of indicated molecules on NP-specific CD8⁺ T cells in the LI/LP and airways of mice shown in C at day 30 PI. (E) Numbers of NP-specific CD8⁺ T cells in each tissue at day 33 and 44 PI (day 3 and 14 after injection). (F) Expression of indicated molecules on NP-specific CD8⁺ T cells in the LI/LP and airways of mice shown in G at day 44 PI. (G) Uninfected mice and mice infected i.p. with x31 were injected i.n. with CpG ODN + NP_366–374 peptide 8 d later as shown in C. Representative dot plots showing the binding of NP_366–374/D^b tetramers on CD8⁺ T cells in the LI/LP mice at day 30 PI (day 22 after injection). (H) Fucci Tg mice were infected i.p. with x31 and then injected i.n. with CpG ODN or CpG ODN + NP_366–374 peptide at day 8 PI. A group of x31-infected mice was challenged i.n. with PR8 at day 30 PI as a positive control to show the proliferating cells in the MLN. Expression of the Cdt1 and Geminin in NP-specific CD8⁺ T cells in the MLN of Fucci Tg mice at day 4 after treatment is shown. Numbers indicate the percentage of cells in each gate. Data are representative of two independent experiments (mean and SEM of five to six mice per group). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 by Student’s t test (A and B) and one-way ANOVA with Tukey’s posthoc tests (C and E).