FIG. 4.

G-specific CD4 T-cell response and lung eosinophilia in rAd/3xG-immune mice. (A) Mice were immunized with rAd i.n. or with vvG by scarification and challenged with RSV 3 weeks after immunization. Lung mononuclear cells were prepared from the lungs of the same groups of mice (n = 4) 4 days after challenge and stimulated with G(183-195) peptide in the presence of brefeldin A. Cells were stained for CD4, IFN-γ, and T-cell receptor Vβ14 and analyzed by flow cytometry. Cells gated for CD4 expression are shown in each dot plot, and the percentages represent the frequency of G-specific IFN-γ-positive cells. The numbers in parentheses indicate the percentage of total Vβ14⁺ cells among CD4⁺ cells. The results are representative of two independent experiments. (B) Lung mononuclear cells were stimulated in vitro with plate-coated anti-CD3 for 48 h, and the levels of IL-4 in the culture supernatant were determined by sandwich ELISA. One representative from two separate experiments is shown. (C) BAL was performed 5 days after challenge, cell pellets were differentially stained, and the number of eosinophils was counted. Data represent means ± standard deviations (n = 4). The results are representative of three independent experiments.