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. 2014 Mar 25;22(7):1364–1374. doi: 10.1038/mt.2014.33

Figure 5.

Figure 5

M2e5x VLP-supplemented vaccination induces IFN-γ producing cellular immune responses. Splenocytes and lung cells were isolated from mice at day 4 postchallenge. IFN-γ secreting cells were determined in the presence of A/California /04/09 (H1N1) viral antigen, M2e peptide, or M2e5x VLPs as a stimulator (2 µg/ml). (a) IFN-γ in splenocytes. (b) IFN-γ in lung cells. IFN-γ cytokine-producing cells were counted by enzyme-linked immunosorbent spot (ELISPOT) reader. P-value indicates significant difference between split+M2e5x and split vaccinated groups. At 4 weeks after boost immunization with M2e5x VLP-supplemented split vaccine, groups of four female Balb/c mice were treated with anti-CD4, anti-CD8, anti-CD4/CD8 monoclonal antibodies or PBS as a control on −2 day and +1 day of challenge. All groups were challenged with a lethal dose of A/Philippines/2/82 (H3N2) influenza virus. (c) Average body weight changes. (d) Survival rates. One asterisk indicates significant difference between PBS and CD4, CD8, or CD4/CD8 T-cell–depleted groups. Two asterisks indicate significant difference between PBS and CD4 or CD4/CD8 T-cell–depleted groups. Three asterisks indicate significant difference between CD4 and CD4/CD8 T-cell–depleted groups. Data represent mean ± SEM. IFN, interferon; M2e, M2 ectodomain; PBS, phosphate-buffered saline; VLP, virus-like particles.