Article Figures & Data
Figures
- Fig 1
PUUV DNA vaccine testing in nonhuman primates. (A) Neutralization of PUUV strain K27. Groups of three rhesus macaques were vaccinated (indicated by arrows) with either pWRG/PUU-M(s1), pWRG/PUU-M(s2), or pWRG/PUU-M(x22). Sera collected on the indicated weeks were tested for the capacity to neutralize PUUV strain K27. Each symbol represents the GMT for 2 or 3 PRNT50 assays. (B) Cross-neutralization of PUUV strain Sotkamo. Sera collected after 3 (week 5) or 4 (week 19) vaccinations were tested by a PRNT using PUUV strain Sotkamo. The PRNT limit of detection was a titer of 20 (dashed line).
- Fig 2
In vivo stability testing of HTNV and PUUV DNA vaccines delivered using a handheld single-use gene gun (ND10). (A) Lot 1 of each vaccine was tested for immunogenicity over 1 year. (B) Lot 2 of each vaccine was tested for immunogenicity over 2 years. Symbols represent the neutralizing antibody PRNT50 titers for individual hamsters vaccinated with either pWRG/HTN-M(x) or pWRG/PUU-M(s2). Hamsters vaccinated with the HTNV DNA vaccine were evaluated for HTNV neutralizing antibodies, and hamsters vaccinated with the PUUV DNA vaccine were evaluated for PUUV strain K27 neutralizing antibodies. The PRNT limit of detection was a titer of 20 (dashed line).
- Fig 3
Evaluation of protection (PUUV) and cross-protection (DOBV) in hamster infection models. Groups of 6 to 8 hamsters were vaccinated (week 0, 2, or 3) with PUUV DNA vaccine, HTNV DNA vaccine, or no vaccine and then challenged with PUUV strain K27 or DOBV. (A) Hamsters were challenged either 4 weeks or 12 weeks after the last vaccination, using an intramuscular challenge of 2,000 PFU PUUV. (B) Hamsters were challenged 11 weeks after the last vaccination, using an intranasal challenge of 2,000 PFU DOBV. Sera were collected at 35 days postchallenge, and N-ELISA log10 titers were plotted. The N-ELISA limit of detection was a titer of 2 log10 (dashed line).
- Fig 4
PUUV DNA vaccine protects against lethal hantavirus disease. Groups of 8 to 10 hamsters were vaccinated with the indicated vaccine or not vaccinated. Sixteen weeks (A) or 20 weeks (B) elapsed before an intranasal challenge of 4,000 PFU ANDV. (C) Groups of 9 or 10 hamsters were vaccinated with the indicated vaccine or not vaccinated before an intramuscular challenge of 2,000 PFU ANDV. Survival of all animals was monitored for 35 days postchallenge. ns, not significant.
Tables
- Table 1
Comparison of differences in deduced M segment open reading frame amino acids for four strains of PUUV and three DNA vaccine constructsf
Amino acid positiona PUUV M segment amino acid DNA vaccine amino acid DTK/Ufa-97b K27c P360d Hallnas B1e PUU-M(x22) PUU-M(s1) PUU-M(s2) 10 Y Y C Y Y Y Y 38 V I I I I I I 187 Q Q Q D Q Q Q 207 H H H H Y Y H 312 A A A V A A A 416 L S L L L L L 448 M M M M T T M 492 W W W W R R W 501 F L F F F F F 604 T T T R T T T 649 L L L H L L L 650 E E E H E E E 675 G G G R G G G 690 P P P Q P P P 972 Q Q Q Q R R Q 1077 G G G P G G G 1097 S S S S L L S - Table 2
PUUV neutralizing antibodies and protection against PUUV infection
Vaccinea Hamster ID Prechallenge PUUV PRNT50 titerd Postchallenge N-ELISA titer (log10)b,d Protectionc pWRG/PUU-M(s2) 269 5,120 3 N 262 2,560 1 Y 261, 263 1,280 1 Y 243 640 1 Y 248 320 2 N 266 320 1 Y 245, 264, 267 160 1 Y 242, 244, 249, 270 80 1 Y 241 40 1 Y 246, 250, 268 20 1 Y 265 1 2 N None 298 40 3 N 281, 284 20 3 N 282 20 2 N 283 1 2 N 285, 286 1 3 N 287 1 4 N 288, 290, 978, 980 1 3 N 291 1 2 N 292, 293–295, 297 1 3 N pWRG/HTN-M(x) 251, 252, 254, 258–260, 232, 236, 239, 240 1 3 N 253, 256, 231, 237 1 4 N ↵a Hamsters were vaccinated on weeks 0, 2, and 3 by use of ND10 delivery devices.
↵b Log10 endpoint titers represent GMTs for two N-ELISAs.
↵c A postchallenge N-ELISA titer of <2 indicates that the hamster was protected. N, not protected; Y, protected.
↵d A value of 1 indicates that the titer was below the limit of detection. The limit of detection for the PRNT was 20, and the limit of detection for the ELISA was 2 (log10).
- Table 3
Calculation of estimated probabilities of protective immunity against PUUV infection based on the protection data from Table 2
Estimated probability of protective immunitya Prechallenge PUUV PRNT50 titer Lower 95% fiducial limit Upper 95% fiducial limit 0.40 24 8 77 0.50 37 13 148 0.60 59 22 305 0.70 97 35 713 0.80 177 58 2,122 0.90 438 119 11,700 ↵a Determined by logistic regression by Probit. Protection status by titer was used to calculate estimated probabilities of protective immunity.
Supplemental material
Files in this Data Supplement:
- Supplemental file 1 -
Fig. S1. Presynthetic PUUV full-length M gene-based DNA vaccine.
PDF, 309K
- Supplemental file 1 -
