Article Figures & Data
Figures
- Fig. 1.
Chemokine mRNA expression in lung tissue of RSV-infected BALB/c mice. Expression of nine murine chemokine mRNAs was investigated by RPA. The figure is a representative result from three independent experiments (six animals per group per experiment). (A) BALB/c mice were infected i.n. with 5 × 105 (lane 4), 106 (lane 3), or 107 (lane 2) PFU of sucrose gradient-purified RSV, or they were sham-infected with uninfected tissue culture medium (lane 5). At day 1 and 5 after infection, RNA was isolated from lung tissue and hybridized with a 32P-labeled RiboQuant MultiProbe (Pharmigen) containing DNA templates for the mouse chemokines Ltn, RANTES, eotaxin, MIP-1β, MIP-1α, MIP-2, IP-10, MCP-1, TCA-3, and the house keeping genes coding for L32 (ribosomal RNA) and GAPDH. After RNAse treatment and purification, protected probes were run on a QuickPoint sequence gel, exposed to an XAR film and developed. The identity of each protected fragment was established as described in Materials and Methods. (B) The quantity of each mRNA species in the original RNA sample was determined based on the signal intensity (by optical densitometry) given by the appropriately sized, protected probe fragment band. Sample loading was normalized to the housekeeping gene GAPDH, included in each template set. The density of each chemokine mRNA on day 1 and day 5 is expressed relative to that of GAPDH.
- Fig. 2.
Chemokine production in lung tissue of RSV-infected mice. Concentrations of RANTES, MIP-1α, and MCP-1 were determined by ELISA in lung tissue homogenate obtained from groups of sham- or RSV-infected mice (24 h). Concentrations of chemokines were adjusted for lung weight. Data are expressed as means + standard errors of the mean (error bars) of three animals per group in three independent experiments. ∗, P < 0.05 in comparison to sham-infected mice; #, P < 0.05 compared with mice infected with 5 × 105 PFU of pRSV; +, P < 0.05 in comparison to mice infected with 106 PFU of pRSV.
- Fig. 3.
Immunolocalization of MIP-1α in lung sections from RSV-infected mice. Frozen lung sections were prepared from 24-h-sham- (A) and -RSV-infected (107 PFU) (B and C) BALB/c mice. MIP-1α was detected by specific polyclonal goat antibody and visualized by Alexa Fluor 488-labeled donkey anti-goat IgG antibody. (A) Sham-infected mice show no evidence of specific staining (magnification, ×50). (B and C) In RSV-infected mice, bright fluorescence staining for MIP-1α appears to be localized in some alveolar epithelial cells and in epithelial cells of some bronchioles and in adjoining capillary endothelium (original magnifications, ×50 [B] and ×10 [C]). The arrows indicate positively stained epithelial cells of bronchioles (BE), endothelial cells (EC), and alveolar epithelial cells (AC).
- Fig. 4.
Lung histopathology of BALB/c mice infected with RSV. Mice were infected i.n. with 5 × 105 PFU (A), 106 PFU (B), and 107 PFU (C) of sucrose-purified RSV or were sham infected (D). At day 1 postinfection mice were killed and lungs were removed, fixed in 10% buffered formalin, and embedded in paraffin. Multiple 4-μm-thick sections were stained with H&E. Original magnification, ×50.
- Fig. 5.
Pathology scores in RSV-infected BALB/c mice. Mice were infected i.n. with 5 × 105, 106, and 107 PFU of sucrose-purified RSV or were sham infected. At day 1 (white bars) and 5 (black bars) postinfection multiple lung sections were stained with H&E. Inflammation was scored blindly by two independent investigators. The number of abnormal perivascular and peribronchial spaces divided by the total spaces counted is the percentage reported as the pathology score (see Materials and Methods). The figure presents the data of two independent experiments with total of five mice per group (mean + standard error of the mean [error bars]).
- Fig. 6.
Viral replication in lung of BALB/c mice infected with sucrose-purified RSV. BALB/c mice (five animals per group) were infected i.n. with 5 × 105, 106, or 107 PFU of RSV (indicated as inoculum). One, five, and twenty-one days after infection, lung tissue was removed and homogenized and the concentration of infectious virus was determined by plaque assay. The mean log10 titer (PFU) per gram of tissue + standard error of the mean (error bar) is shown. The lower detection limit is indicated.
- Fig. 7.
Chemokine mRNA expression in lung of mice inoculated with UV-inactivated RSV. BALB/c mice were inoculated i.n. with UV-inactivated purified RSV (107 PFU) (lane UV), or they were sham-infected (lane S). At day 1 after infection, chemokine mRNA expression was determined by RPA as described in the legend to Fig. 1. The figure is a representative result from three animals per group.
- Fig. 8.
Chemokine mRNA expression in lung tissue of RSV-infected MIP-1α−/− mice. Expression of chemokine mRNA was determined exactly as described in the legend to Fig. 1. MIP-1α−/− mice (−/−) and control littermates (+/+) were infected i.n. with 107 PFU of purified RSV, or they were sham inoculated. At day 5 after infection, extracted lung RNA was analyzed by RPA. The density of each chemokine mRNA in RSV-infected MIP-1α−/− and MIP-1α+/+ mice is expressed relative to that of GAPDH. The data are expressed as means + standard errors of the means (error bars) of four animals per group.
- Fig. 9.
Lung histopathology of RSV-infected MIP-1α−/− mice. MIP-1α−/− mice and control littermates were infected i.n. with 107 PFU of sucrose gradient-purified RSV or were sham inoculated. At day 5 postinfection lung sections were stained with H&E and inflammation was scored as described in Materials and Methods. A representative section is shown from a +/+ control mouse (A) and from a −/− mouse (B). Original magnification, ×50.
