TABLE 1.
Virusa | Plaque sizeb | Temp sensitivity (log10 no. of PFU/ml)
|
Mouse neuroinvasiveness
|
|||
---|---|---|---|---|---|---|
37.0°C | 41.0°C | Δc | i.p. LD50 (PFU) | AST ± SDd | ||
NY99 (wt virus) | lp | 8.1 | 7.6 | 0.5 | 0.8 | 8.0 ± 1.2 |
NY99ic | lp | 7.0 | 7.3 | 0.3 | 1.0 | 7.2 ± 0.6 |
Bird 1153 (wt virus) | sp | 7.7 | 4.8 | 2.9† | >10,000e | NA |
NS4B-E249G mutant | lp | 6.2 | 6.0 | 0.2 | 1.2 | 7.5 ± 0.4 |
prM-V156I mutant | lp | 5.9 | 6.1 | 0.2 | 0.7 | 8.6 ± 1.4 |
NS5-A804V mutant | mp | 6.3 | 5.5 | 0.8 | 5 | 8.0 ± 0.5 |
3′UTRB mutant | lp | 6.4 | 5.0 | 1.4* | 0.6 | 8.8 ± 2.0 |
prM-V156I + NS4B-E249G mutant | lp | 7.0 | 7.2 | 0.2 | 1.4 | 8.8 ± 1.5 |
prM-V156I + 3′UTRB mutant | lp | 6.6 | 5.1 | 1.5* | 4.2 | 9.0 ± 1.0 |
NS4B-E249G + 3′UTRB mutant | sp | 6.7 | 3.9 | 2.8† | >10,000e | NA |
NS4B-E249G + NS5-A804V mutant | sp | 6.2 | 6.4 | 0.2 | 2,000e | 9.2 ± 1.2 |
NS5-A804V + 3′UTRB mutant | mp | 5.4 | 4.2 | 1.2* | >10,000e | NA |
The wt NY99 and Bird 1153 WNV isolates are indicated. The remaining mutant-virus designations indicate each Bird 1153 virus-specific locus incorporated into the NY99ic viral background.
Plaque sizes were as follows: sp were <1.0 mm, lp were >3.0 mm, and mp were 1 to 3 mm in Vero cells.
Δ indicates the difference between average virus titers at 37.0°C and 41.0°C (triplicate experiments) at 72 h postinfection in Vero cells. † indicates a P of <0.001 in a comparison of the viral titer at 41.0°C versus that at 37.0°C, with experiments performed in triplicate. * indicates a P of <0.05 in a comparison of the viral titer at 41.0°C versus that at 37.0°C, with experiments performed in triplicate.
AST, average survival time ± standard deviation following intraperitoneal inoculation of virus. NA, not applicable, as no mice died following inoculation.
Attenuated for mouse neuroinvasiveness.