FIG 4.
CD8 T cell response induced by X31-sciIV. Female C57BL/6 mice were i.n. inoculated with 105 PFU of X31-sciIV virus, 105 PFU of WT X31 virus, or PBS control. At 10 days after inoculation, the spleens, mediastinal lymph nodes (MLN), lungs, and bronchoalveolar lavage (BAL) tissues were harvested for lymphocyte preparation and FACS staining. (A) Total CD8 T cells in the lungs and BAL fluid. The total number of lung/BAL CD8 T cells was obtained by multiplying the total number of lung or BAL lymphocytes counted by the percentage of CD8 T cells in the total gated lymphocyte population. The data represent averages ± the SD. The differences in the total CD8 T cells in the lungs or BAL fluid among three groups of animals were compared and analyzed by using one-way ANOVA. (B) Tetramer analysis of influenza virus-specific CD8 T cells in the lungs and BAL fluid. Lung or BAL single-cell suspensions were stained with Live/Dead dye, anti-CD3, anti-CD8 antibodies, and H2Db/NP366 and H2Db/PA224 tetramers. The data are representative of three independent experiments. (C) Total NP366- and PA224-specific CD8 T cells in lungs and BAL fluid. The total number of lung or BAL CD8 T cells was obtained by multiplying the total number of lung or BAL CD8 T cells by the percentage of tetramer-positive cells. The data represent averages ± the SD. The differences in the influenza virus NP366- or PA224-specific CD8 T cells in the lungs or BAL fluid among three groups of mice were compared and analyzed by using one-way ANOVA. (D) Tetramer analysis of influenza virus-specific CD8 T cells in lymphoid tissues. At day 10 after inoculation, the spleens and MLNs were collected for single-cell suspension preparation and staining with Live/Dead dye, anti-CD3, anti-CD8 antibodies, and H2Db/NP366 and H2Db/PA224 tetramers. The data are representative of three independent experiments.