Figure 6

Assessment of susceptibility of RA, CS, HK-NTHi or CS/HK-NTHi-exposed mice to RV infection. RA, CS, HK-NTHi or CS/HK-NTHi-exposed mice were either infected with RV or equal volume of sham and mice were sacrificed 4 days post-infection. (A) Total RNA was isolated from whole lungs and viral RNA (vRNA) copy number was determined by quantitative qPCR and expressed vRNA copy number per 10 μg of total RNA. (B) Lung homogenates were used to determine infectious viral load by assessing number of plaque forming units (PFU) per lung. (C and D). Cells from bronchoalveolar lavage were used to determine number of neutrophils and lymphocytes and expressed as cells/ml of BAL. Data in A to D represent median with range calculated from 5 to 6 mice per group (* different from RA-exposed mice, p ≤ 0.05, ANOVA on ranks; # different from CS-exposed mice, p ≤ 0.05, ANOVA on ranks; † different from HK-NTHi-exposed mice, p ≤ 0.05, ANOVA on ranks). (E and F) Supernatants from bronchoalveolar lavage were used to determine KC and MIP2 levels. (G) Total lung RNA was reverse transcribed and subjected to qPCR to determine mRNA levels of IP-10 and expressed as fold change over house-keeping gene, G3PDH. Data in E to G represents mean ± SD calculated from 5–6 mice per group (* different from respective sham controls, p ≤ 0.05, ANOVA; # different from CS-exposed RV-infected mice, p ≤ 0.05, ANOVA; † different from HK-NTHi-exposed RV-infected mice, p ≤ 0.05, ANOVA).