Survival and bacterial burden of mice infected with wild-type, pneumolysin gene (ply)–deleted, and ply-recoded Streptococcus pneumoniae serotype 3 (SP3) strains. A. Survival of BALB/c mice after intranasal infection with 5 × 104 colony-forming units (CFUs) of wild-type, ply-recoded, and ply-deleted SP3 A66.1 strains. **P < .001 comparing A66.PM4 and A66.1; **P < .002 comparing A66:PM4 and A66:Δply; *P < .02 comparing A66:PM2 and A66:Δply; *P < .001 comparing A66:PM2 and A66.1 (Kaplan-Meier log-rank test; curves depict results of 2 independent experiments with 5 mice per group; n = 10). B, CFUs in lungs of mice infected as in panel A, 48 h after infection. *P < .05 comparing A66.1 and A66:PM4; *P < .05 comparing A66.1 and A66:Δply (Student unpaired t test). C, CFUs in blood of mice infected as in panel A, 48 h after infection. No S. pneumoniae was detected in the blood of mice infected with A66:PM4 (lower limit of detection, 101). For panels B and C, bars represent the mean and error bars represent the standard deviation (n = 3).