Ag 85A–specific immune responses.

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Ag 85A–specific immune responses. Ag 85A–specific immune responses. (A) Mice were infected with eMCMV or MCMV85A and 4 wk later boosted with 2 × 109 virus particles Ad85A i.m. Six days later, lung, spleen, and liver cells were isolated and stimulated for 6 h with pooled 85A peptides. (B) Spleen cells from eMCMV- or MCMV85A-infected mice were stimulated in vitro with a pool of 15-mer peptides covering the sequence of Ag 85A, expanded with IL-2, rested, and restimulated with 85A peptides before assaying by intracellular staining and flow cytometry for Ag-specific IFN-γ–producing cells. (C) Lung or spleen cells from MCMV85A-infected mice boosted with Ad85A were stimulated with individual 85A peptides covering the dominant H-2d CD4 and CD8 epitopes. The frequencies of IFN-γ– or IL-2–producing cells were determined by flow cytometry on CD8- and CD4-gated cells. Results are expressed as the means ± SD of three or four mice per group, representative of two independent experiments. Data were analyzed by a one-way ANOVA with a Tukey posttest. Peter C. L. Beverley et al. J Immunol 2014;193:2306-2316 Copyright © 2014 The Authors