Fig 1.
Schematic of genome organization of wild-type VSV (top), rVSV-EBOV (Ebola) vector (center) showing the substitution of the EBOV GP gene for the glycoprotein (G) gene and principal virulence factor of wild-type VSV, and rVSV-Nipah (bottom) showing the addition of NiV G encoding the NiV attachment glycoprotein downstream of the EBOV GP.
Fig 2.
Survival distribution, infant mice and hamsters inoculated by the IC route.
A. 8-day-old ICR mice. Twenty μL of PHV02 at graded doses or YF 1DD vaccine were inoculated into 2 litters (10 suckling pups/litter). The doses injected in 20 μL were 100,000, 1000, or 10 pfu for PHV02 groups and 640 pfu for YF 17DD. Animals were observed for 21 consecutive days and clinical signs and deaths recorded. All mice died in the intermediate and high dose groups, 85% died in the low dose (10 pfu) group and 70% died in the reference group receiving 640 pfu of YF17DD. B. 8-day-old Golden hamsters. Twenty μL of PHV02 at graded doses or YF 17DD were inoculated into 2 litters (8-10 suckling pups/litter). The doses injected in 20 μL were 1000 or 10 pfu for PHV02 groups and 640 pfu for YF 17DD. All infant hamsters died, but the YF 17DD animals had a statistically longer survival time (see text). Since all animals in the test article groups died by Day 7, the study was terminated at that point.
Fig 3.
Survival distribution, 6- to 8-week-old ICR mice and Golden hamsters inoculated by the IC route.
6- to 8-week-old ICR mice Twenty μL of PHV02 at graded doses or YF 17DD were inoculated IC into groups of 5 mice. The doses injected in 20 μL were 100,000, 1000, or 10 pfu for PHV02 groups and 640 pfu for YF 17DD. 6- to 8-week-old Golden hamsters. Twenty μL of PHV02 or YF 17DD were inoculated IC into groups of 5 hamsters. The doses injected in 20 μL were 100,000, 1000, or 10 pfu for PHV02 groups and 640 pfu for YF 17DD. Animals were observed for 21 consecutive days and clinical signs and deaths recorded. Hamsters inoculated with 100,000 pfu of PHV02 virus had a 60% mortality ratio for an IC LD50 of 46,800 pfu. In contrast to adult mice which are susceptible to lethal IC infection with YF 17DD Fig 3A), adult hamsters showed no illness or deaths.
Fig 4.
Survival distribution of infant and adult Swiss-Webster mice inoculated IC with rVSV-Nipah or rVSV-EBOV.
A. 8-day-old mice (10 animals/group) inoculated IC with 0.1, 1, 10 or 100 pfu of PHV02 (VSV-Nipah virus), rVSV-EBOV, or 0.9% saline. rVSV-EBOV was avirulent at all dose levels. The IC LD50 for PHV02 is 5.3 pfu, and the LD50 of rVSV-EBOV is >100 pfu. B. 6- to 8-week-old Swiss-Webster mice (10 animals/group) inoculated IC with 100,000 pfu of VSV-Nipah virus, rVSV-EBOV, or 0.9% saline.
Table 1.
Design of the Monkey Neurovirulence Test.
Table 2.
Monkey Neurovirulence Test: Individual Brain and Spinal Cord Scoresa, Cynomolgus Macaques, evaluated 30 days after inoculation.
Table 3.
Discriminator, Target, and Combined Scores, Cynomolgus Macaques, Neurovirulence Test, evaluated 30 days after inoculation.