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. 2016 Jul 7;1(10):e85832. doi: 10.1172/jci.insight.85832

Figure 3. LAIV-vaccination generates lung TRM.

Figure 3

(A) Lung T cells labeled by i.v. anti-Thy1 antibody in mice 6 weeks after vaccination with 2014–2015 IIV or LAIV. Left: Representative flow cytometry plots displaying CD4+ and CD8+ cells in tissues (“Protected”) or circulating (“Labeled”). Right: Individual percentages of protected T cells ± SEM (n = 5 mice per group, representative of 2 experiments; significance determined by 2-way ANOVA with Holm-Sidak’s multiple comparisons test, ****P < 0.0001, ***P < 0.001, *P < 0.05). (B) Protected lung CD69+ T cells in mice 16 weeks after vaccination with 2014–2015 IIV or LAIV or after infection with PR8 influenza. Graph displays protected CD69+ cell numbers ± SEM (n = 4–5 mice per group, representative of 2 experiments; significance determined by multiple Student’s t tests with Welch’s correction, *P < 0.05). (C) CD69 expression by protected versus circulating lung T cells 16 weeks after 2014–2015 LAIV vaccination or PR8 infection. Top: Representative histogram of CD69 expression by protected/labeled lung T cells after PR8 infection. Bottom: CD69 expression by protected/labeled lung T cells after LAIV vaccination. (D) TRM marker expression in protected lung T cells 6 weeks after vaccination with 2015–2016 IIV or LAIV. Left: Representative flow cytometry plots displaying protected CD4+ and CD8+ T cells with CD44hiCD69+ phenotypes or protected CD8+CD44+ T cells with CD69+CD103hi phenotype. Right: Individual percentages of protected CD4+ or CD8+ T cells expressing CD69 or protected CD8+CD44+ T cells expressing CD103 ± SEM (n = 5 mice per group, representative of 3 experiments; significance determined by multiple Student’s t tests with Holm-Sidak’s correction, ****P < 0.0001, ***P < 0.001). (E) Protected lung influenza–specific CD8+ T cells in mice 5 weeks after vaccination with 2015–2016 IIV or LAIV. Left: Representative flow cytometry plots displaying protected or labeled lung NP366-374–specific CD8+ T cells. Right: Individual percentages of protected lung NP366-374–specific CD8+ T cells ± SEM (n = 3–5 mice per group; significance determined by 1-way ANOVA with Holm-Sidak’s multiple comparisons test, ***P < 0.001).