FIG. 2.

Validation of semiquantitative RT-PCR assays to study chemokine gene expression profiles. Eotaxin, MCP-1, and MIP-2 protein and gene expression profiles were compared in mouse lung tissue extracts following RSV challenge, as a function of priming antigen. Groups of six mice were immunized three times i.p. with 20 or 50 μg BBG2Na, FI-RSV (stock 2 diluted at 1/100), or PBS in the presence of Alhydrogel. Mice were sacrificed 5 days after i.n. challenge with RSV. Lungs were removed and processed for either relative chemokine protein (solid bars) or gene expression (open bars) levels, as described in Materials and Methods. Chemokine protein concentrations are presented as fractions of total lung homogenate protein concentrations, i.e., relative chemokine protein concentration = [chemokine] (pg/ml)/[total protein] (mg/ml). Chemokine gene expression data are presented as means + SD (error bars) of OD₄₀₅ for the chemokine mRNA relative to the OD₄₀₅ for the β-actin mRNA.