A Non-Hemadsorbing Live-Attenuated Virus Vaccine Candidate Protects Pigs against the Contemporary Pandemic Genotype II African Swine Fever Virus
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cells and Viruses
2.2. Animals
2.3. Virus Passage, Titration, and Replication Test In Vitro
2.4. ASFV Genome Next-Generation Sequencing
2.5. Safety, Efficacy, and Duration of Protection Testing of VNUA-ASFV-LAVL3 in Pigs
2.6. Quantitative PCR (qPCR) for ASFV
2.7. Detection of ASFV-Specific Antibody and Cellular Responses in Pigs
2.8. Statistical Analysis
3. Results
3.1. VNUA-ASFV-LAVL3 Loses Hemadsorption Activity during Adaptation in 3D4/21 Cells
3.2. Genetic Characterization of VNUA-ASFV-LAVL3
3.3. VNUA-ASFV-LAVL3 Exhibits High Safety in Pigs
3.4. Vaccination with VNUA-ASFV-LAVL3 Confers Full Protection in Pigs
3.5. VNUA-ASFV-LAVL3 Vaccination Prevents Replication of Wild-Type ASFV and Protects against Pathological Lesions in Pigs
3.6. Pigs with VNUA-ASFV-LAVL3 Induce Robust ASFV-Specific Humoral and Cellular Immune Responses
3.7. Pigs Can Be Protected against Wild-Type ASFV Challenge for Up to 2 Months after a Single Vaccination with VNUA-ASFV-LAVL3
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Genome Position | Mutation Type | Gene | Nucleotide Change | Amino Acid Change | Protein Change |
---|---|---|---|---|---|
6343 to 6344 | Insertion | Uncharacterized | 61nt | Uncharacterized | Uncharacterized |
6366 | Substitution | Uncharacterized | T to A | Uncharacterized | Uncharacterized |
6371 | Substitution | Uncharacterized | G to A | Uncharacterized | Uncharacterized |
8555 to 8718 | Deletion | Uncharacterized | Deletion | Deletion | Deletion |
8719 to 9003 | Deletion | 285L | Deletion | Deletion | Deletion |
9004 to 9131 | Deletion | Uncharacterized | Deletion | Deletion | Deletion |
9132 to 9515 | Deletion | MGF110-8L | Deletion | Deletion | Deletion |
9516 to 9732 | Deletion | Uncharacterized | Deletion | Deletion | Deletion |
9733 to 10107 | Deletion | MGF_100-1R | Deletion | Deletion | Deletion |
10108 to 10265 | Deletion | Uncharacterized | Deletion | Deletion | Deletion |
10266 to 11138 | Deletion | MGF_100-9L | Deletion | Deletion | Deletion |
11139 to 11428 | Deletion | Uncharacterized | Deletion | Deletion | Deletion |
11429 to 11788 | Deletion | MGF110-11L | Deletion | Deletion | Deletion |
11789 to 11877 | Deletion | Uncharacterized | Deletion | Deletion | Deletion |
11878 to 12243 | Deletion | MGF110-14L | Deletion | Deletion | Deletion |
12244 to 12432 | Deletion | Uncharacterized | Deletion | Deletion | Deletion |
12433 to 12792 | Deletion | MGF110-12L | Deletion | Deletion | Deletion |
12793 to 12878 | Deletion | Uncharacterized | Deletion | Deletion | Deletion |
12879 to 13702 | Deletion | MGF110-13L | Deletion | Deletion | Deletion |
13703 to 13882 | Deletion | Uncharacterized | Deletion | Deletion | Deletion |
13883 to 15046 | Deletion | MGF360-4L | Deletion | Deletion | Deletion |
15047 to 15862 | Deletion | Uncharacterized | Deletion | Deletion | Deletion |
15863 to 16990 | Deletion | MGF360-6L | Deletion | Deletion | Deletion |
16991 to 17868 | Deletion | Uncharacterized | Deletion | Deletion | Deletion |
17869 to 17972 | Deletion | X69R | 104 nt | 34 aa | Shorter protein |
20076 | Deletion | MGF_300-2R | C | D to I | Disruption |
20188 | Substitution | MGF_300-2R | T to C | S to N | Disruption |
23952–23962 | Deletion | Uncharacterized | 11 nt | Uncharacterized | Uncharacterized |
24103 | Substitution | MGF_360-10L | T to C | D to Q | Disruption |
24752 | Deletion | MGF_360-10L | A | D to E | Disruption |
28221–28324 | Deletion | MGF_360-12L | 104 nt | 34aa | Shorter protein |
30324 | Deletion | Uncharacterized | A | Uncharacterized | Uncharacterized |
30871 | Substitution | MGF_360-14L | G to A | H to A | Abnormal protein |
30876–30877 | Substitution | MGF_360-14L | TC to AT | I to S | Abnormal protein |
30879 | Substitution | MGF_360-14L | C to T | D to V | Abnormal protein |
30882–30915 | Deletion | MGF_360-14L | 34 nt | 11 aa | Abnormal protein |
30918 | Substitution | MGF_360-14L | T to A | N to Y | Abnormal protein |
30923 | Substitution | MGF_360-14L | T to A | Y to F | Abnormal protein |
30930 | Substitution | MGF_360-14L | T to A | S to C | Abnormal protein |
30938 | Substitution | MGF_360-14L | A to C | V to G | Abnormal protein |
33291 | Deletion | MGF_505-2R | A | K to T | Shorter protein |
36919 | Substitution | MGF_505-5R | T to C | Y to N | Substitution |
40351 | Substitution | MGF_505-7R | T to G | V to G | Substitution |
41114 | Deletion | Uncharacterized | T | Uncharacterized | Uncharacterized |
42931–42947 | Deletion | Uncharacterized | 17 nt | Uncharacterized | Uncharacterized |
43100–43133 | Deletion | Uncharacterized | 34 nt | Uncharacterized | Uncharacterized |
46011 | Deletion | A104R | A | K to S | Disruption |
53169 | Deletion | Uncharacterized | T | Uncharacterized | Uncharacterized |
58635 | Substitution | F1055L | T to C | I to W | Disruption |
58913 | Substitution | F1055L | T to G | H to L | Disruption |
60945–61020 | Deletion | F1055L | 76 nt | 25 aa | Disruption |
61819 | Substitution | Uncharacterized | T to C | Uncharacterized | Uncharacterized |
72222 | Substitution | EP402R | T to C | C to R | Disruption |
72824 | Deletion | EP402R | T | R to E | Disruption |
72965 | Deletion | EP402R | C | P to T | Disruption |
75240–75320 | Deletion | M1249L | 81 nt | 27 aa | Abnormal protein |
77036 | Substitution | M1249L | T to C | M to V | Abnormal protein |
78038 | Substitution | M1249L | T to G | N to H | Abnormal protein |
85588 | Substitution | C475L | T to G | R to S | Substitution |
93313 | Substitution | B962L | A to G | Y to H | Substitution |
102005 | Substitution | B385R | A to G | T to A | Substitution |
105743–105744 | Insertion | B407L | 40nt | 13 aa | Disruption |
105777 | Substitution | B407L | T to C | S to G | Disruption |
107968–107969 | Insertion | G1340L | 19 nt | 6 aa | Abnormal protein |
109399 | Substitution | G1340L | A to C | V to G | Abnormal protein |
114357–114561 | Deletion | G1211R | 205 nt | 68 aa | Disruption |
120939–121034 | Deletion | CP2475L | 96 nt | 32 aa | Abnormal protein |
122809 | Substitution | CP2475L | T to C | N to S | Abnormal protein |
124660 | Substitution | CP530R | A to G | N to D | Substitution |
130367–130446 | Deletion | NP1450L | 80 nt | 26 aa | Disruption |
131201–131500 | Deletion | NP1450L | 300 nt | 100 aa | Disruption |
131815 | Substitution | NP1450L | A to G | I to T | Disruption |
136825–136841 | Deletion | D250R | 17 nt | 6 aa | Shorter protein |
141015 | Substitution | D1133L | T to C | E to A | Disruption |
141043 | Substitution | D1133L | T to G | T to S | Disruption |
141285–141420 | Deletion | D1133L | 136 nt | 48 aa | Disruption |
145156–145171 | Deletion | S183L | 16 nt | 5 aa | Disruption |
145490 | Substitution | S273R | C to A | S to S | None |
146353–146375 | Deletion | P1192R | 23 nt | 7 aa | Disruption |
153308 | Substitution | H233R | T to C | V to A | Disruption |
153598–153665 | Deletion | H233R | 68 | 22 aa | Disruption |
154180 | Substitution | H240R | C to T | L to F | Substitution |
166366 | Substitution | E248R | A to C | P to P | None |
173569–173592 | Deletion | I177L | 24 | 8 aa | Abnormal protein |
174320–174320 | Deletion | Uncharacterized | A | Uncharacterized | Uncharacterized |
182192 | Deletion | MGF 360-18R | A | E to N | Disruption |
182665 | Substitution | MGF 360-18R | T to C | L to N | Disruption |
Organs | Quantitative ASFV Real-Time PCR | |||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
103 TCID50 Vaccinated (Pig Number) | 104 TCID50 Vaccinated (Pig Number) | 105 TCID50 Vaccinated (Pig Number) | Control (Pig Number) | |||||||||||||||
80 | 81 | 82 | 83 | 84 | 85 | 86 | 87 | 88 | 89 | 90 | 91 | 92 | 93 | 94 | 96 | 97 | 98 | |
Brain | - | - | - | - | - | - | - | - | - | - | - | - | - | - | - | + | + | + |
Heart | - | - | - | - | - | - | - | - | - | - | - | - | - | - | - | + | + | + |
Lung | - | - | - | - | - | - | - | - | - | - | - | - | - | - | - | + | + | + |
Liver | - | - | - | - | - | - | - | - | - | - | - | - | - | - | - | + | + | + |
Stomach | - | - | - | - | - | - | - | - | - | - | - | - | - | - | - | + | + | + |
Spleen | - | - | - | - | - | - | - | - | - | - | - | - | - | - | - | + | + | + |
Kidney | - | - | - | - | - | - | - | - | - | - | - | - | - | - | - | + | + | + |
Bladder | - | - | - | - | - | - | - | - | - | - | - | - | - | - | - | + | + | + |
Tonsil | - | - | - | - | - | - | - | - | - | - | - | - | - | - | - | + | + | + |
ILN | - | - | - | - | - | - | - | - | - | - | - | - | - | - | - | + | + | + |
MLN | - | - | - | - | - | - | - | - | - | - | - | - | - | - | - | + | + | + |
SLN | - | - | - | - | - | - | - | - | - | - | - | - | - | - | - | + | + | + |
BM | - | - | - | - | - | - | - | - | - | - | - | - | - | - | - | + | + | + |
Group | No. of Pigs | Vaccination | Wild-Type ASFV Challenge at | Mean (±SD) Time to Death (Days) | Mean (±SD) Fever Duration (Days) | Survival/Protection Rate (%) | Clinical Signs |
---|---|---|---|---|---|---|---|
A | 5 | 104 VNUA-ASFV-LAVL3 | 1-month post-vaccination | 0 | 0 | 5/100 | Healthy |
3 | No vaccination, contact | 10.3 (±0.57) | 5.6 (±0.57) | 0/0 | High fever, ASF clinical signs | ||
B | 3 | No vaccination | 8.3 (±0.57) | 4.3 (±0.57) | 0/0 | High fever, ASF clinical signs | |
C | 5 | 104 VNUA-ASFV-LAVL3 | 2-month post-vaccination | 0 | 0 | 5/100 | Healthy |
3 | No vaccination, contact | 10.3 (±0.57) | 5.6 (±0.57) | 0/0 | High fever, ASF clinical signs | ||
D | 3 | No vaccination | 8.6 (±0.57) | 4.6 (±0.57) | 0/0 | High fever, ASF clinical signs |
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© 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Truong, Q.L.; Wang, L.; Nguyen, T.A.; Nguyen, H.T.; Le, A.D.; Nguyen, G.V.; Vu, A.T.; Hoang, P.T.; Le, T.T.; Nguyen, H.T.; et al. A Non-Hemadsorbing Live-Attenuated Virus Vaccine Candidate Protects Pigs against the Contemporary Pandemic Genotype II African Swine Fever Virus. Viruses 2024, 16, 1326. https://doi.org/10.3390/v16081326
Truong QL, Wang L, Nguyen TA, Nguyen HT, Le AD, Nguyen GV, Vu AT, Hoang PT, Le TT, Nguyen HT, et al. A Non-Hemadsorbing Live-Attenuated Virus Vaccine Candidate Protects Pigs against the Contemporary Pandemic Genotype II African Swine Fever Virus. Viruses. 2024; 16(8):1326. https://doi.org/10.3390/v16081326
Chicago/Turabian StyleTruong, Quang Lam, Lihua Wang, Tuan Anh Nguyen, Hoa Thi Nguyen, Anh Dao Le, Giap Van Nguyen, Anh Thi Vu, Phuong Thi Hoang, Trang Thi Le, Huyen Thi Nguyen, and et al. 2024. "A Non-Hemadsorbing Live-Attenuated Virus Vaccine Candidate Protects Pigs against the Contemporary Pandemic Genotype II African Swine Fever Virus" Viruses 16, no. 8: 1326. https://doi.org/10.3390/v16081326