Clin Exp Vaccine Res.  2015 Jan;4(1):114-118. 10.7774/cevr.2015.4.1.114.

Antigenic properties and virulence of foot-and-mouth disease virus rescued from full-length cDNA clone of serotype O, typical vaccine strain

Affiliations
  • 1Animal and Plant Quarantine Agency, Anyang, Korea. parkjhvet@korea.kr
  • 2Clinical Research Center of the Affiliated Hospital of Guangdong Medical College, Zhanjiang, China.

Abstract

We cloned the full-length cDNA of O Manisa, the virus for vaccinating against foot-and-mouth disease. The antigenic properties of the virus recovered from the cDNA were similar to those of the parental virus. Pathogenesis did not appear in the pigs, dairy goats or suckling mice, but neutralizing antibodies were raised 5-6 days after the virus challenge. The utilization of O Manisa as a safe vaccine strain will increase if recombinant viruses can be manipulated by inserting or removing a marker gene for differential serology or replacing the protective gene from another serotype.

Keyword

Foot-and-mouth disease; Vaccine; Molecular cloning

MeSH Terms

Animals
Antibodies, Neutralizing
Clone Cells*
Cloning, Molecular
DNA, Complementary*
Foot-and-Mouth Disease
Foot-and-Mouth Disease Virus*
Goats
Humans
Mice
Parents
Swine
Virulence*
Antibodies, Neutralizing
DNA, Complementary

Figure

  • Fig. 1 Recovery of virus from FMDV full-length cDNA clone, and antibody response, pathogenesis in animals after infection of the virus. (A) Recovery of virus from full-length cDNA clone (pO-Manisa-FG) from foot-and-mouth disease virus. The small fragment (1-369 nt), including poly(C) of O Manisa, were cloned into pBluescript II KS (+) vector. The large (382-8,206 nt) were subsequently inserted into plasmid. The redundant restriction enzyme sites were removed (B). The LF-BK cells were overlaid with 2 mL of 1.5% SeaPlaque agarose containing 2% fetal bovine serum in Dulbecco's modified Eagle's medium and cultured at 37℃ in 5% CO2 for three days. The plaques were visualized by staining with a 0.1% crystal violet solution. The recovered virus had a plaque morphology similar to that of the parental virus. For electron microscopy of O Manisa virus, virion purification was performed by sucrose gradient centrifugation. The viruses were concentrated using Amicon ultracentrifuge filters (100 kDa). Samples were observed using electron microscope. The viruses showed size of 28 nm. (C) Growth properties of FMDV, O Manisa-parental virus and O Manisa-FG recovered from pO-Manisa-FG clone. One-step growth kinetics in BHK21 (up-left) or LF-BK cells (up-right). Virus titration by tissue culture (down-left) and quantitative reverse transcription polymerase chain reaction (down-right). There was no significant difference in growth characterization between transfectant virus and the parental virus. (D) VN antibody level after challenge of O Manisa-FG in the pigs. Pigs were challenged by different administration routes (ID injection on the foot-pad with 0.1 mL, and IM injection 1 mL, with 105.0 TCID50/0.1 mL). The animals did not show clinical signs. The viruses for VN test were O Manisa-parental virus (left) and O/Andong/SKR/2010 strain of SEA topotype (right). (E) VN antibody level after challenge of O Manisa-FG in the dairy goats. Dairy goats were challenged by ID route of the same method as pig injection and the viruses for VN test were O Manisa-parental virus (left) and O/Andong/SKR/2010 (right). (F) Survival in seven-day-old mice after challenge of recovered, parental virus and O/SKR/2002 of 105.0 TCID50/0.1 mL by intraperitoneal route. FMDV, foot-and-mouth disease virus; VN, virus neutralizing; ID, intradermal; IM, intramuscular; SEA, South East Asian.


Reference

1. Elnekave E, Li Y, Zamir L, et al. The field effectiveness of routine and emergency vaccination with an inactivated vaccine against foot and mouth disease. Vaccine. 2013; 31:879–885.
Article
2. Hema M, Chandran D, Nagendrakumar SB, Madhanmohan M, Srinivasan VA. Construction of an infectious cDNA clone of foot-and-mouth disease virus type O 1 BFS 1860 and its use in the preparation of candidate vaccine. J Biosci. 2009; 34:45–58.
Article
3. Knowles NJ, He J, Shang Y, et al. Southeast Asian foot-and-mouth disease viruses in Eastern Asia. Emerg Infect Dis. 2012; 18:499–501.
Article
4. Rajasekhar R, Hosamani M, Basagoudanavar SH, et al. Rescue of infective virus from a genome-length cDNA clone of the FMDV serotype O (IND-R2/75) vaccine strain and its characterization. Res Vet Sci. 2013; 95:291–297.
Article
5. Li P, Bai X, Lu Z, et al. Construction of a full-length infectious cDNA clone of inter-genotypic chimeric foot-and-mouth disease virus. Wei Sheng Wu Xue Bao. 2012; 52:114–119.
6. Lu S, Zhao Q, Liu X, et al. Construction of an infectious cDNA clone derived from foot-and-mouth disease virus O/QYYS/s/06. Sheng Wu Gong Cheng Xue Bao. 2009; 25:982–986.
7. Li P, Bai X, Sun P, et al. Evaluation of a genetically modified foot-and-mouth disease virus vaccine candidate generated by reverse genetics. BMC Vet Res. 2012; 8:57.
Article
8. Chang Y, Zheng H, Shang Y, et al. Recovery of infectious foot-and-mouth disease virus from full-length genomic cDNA clones using an RNA polymerase I system. Acta Biochim Biophys Sin (Shanghai). 2009; 41:998–1007.
Article
9. Liu G, Liu Z, Xie Q, et al. Generation of an infectious cDNA clone of an FMDV strain isolated from swine. Virus Res. 2004; 104:157–164.
Article
10. Zibert A, Maass G, Strebel K, Falk MM, Beck E. Infectious foot-and-mouth disease virus derived from a cloned full-length cDNA. J Virol. 1990; 64:2467–2473.
Article
11. Uddowla S, Hollister J, Pacheco JM, Rodriguez LL, Rieder E. A safe foot-and-mouth disease vaccine platform with two negative markers for differentiating infected from vaccinated animals. J Virol. 2012; 86:11675–11685.
Article
12. Li P, Bai X, Cao Y, et al. Expression and stability of foreign epitopes introduced into 3A nonstructural protein of foot-and-mouth disease virus. PLoS One. 2012; 7:e41486.
Article
13. Ito N, Takayama-Ito M, Yamada K, Hosokawa J, Sugiyama M, Minamoto N. Improved recovery of rabies virus from cloned cDNA using a vaccinia virus-free reverse genetics system. Microbiol Immunol. 2003; 47:613–617.
Article
14. Escarmis C, Toja M, Medina M, Domingo E. Modifications of the 5' untranslated region of foot-and-mouth disease virus after prolonged persistence in cell culture. Virus Res. 1992; 26:113–125.
Article
15. Saiz M, Gomez S, Martinez-Salas E, Sobrino F. Deletion or substitution of the aphthovirus 3' NCR abrogates infectivity and virus replication. J Gen Virol. 2001; 82:93–101.
Article
16. Grubman MJ, Baxt B, Bachrach HL. Foot-and-mouth disease virion RNA: studies on the relation between the length of its 3'-poly(A) segment and infectivity. Virology. 1979; 97:22–31.
Article
Full Text Links
  • CEVR
Actions
Cited
CITED
export Copy
Close
Share
  • Twitter
  • Facebook
Similar articles
Copyright © 2024 by Korean Association of Medical Journal Editors. All rights reserved.     E-mail: koreamed@kamje.or.kr