Expression and characterization of the two outer capsid proteins of African horsesickness virus: the role of VP2 in virus neutralization.
Abstract: African horsesickness virus (AHSV) is a gnat-transmitted member of the Orbivirus genus of the Reoviridae family. The virus has a genome of 10 double-stranded RNA species (L1-L3, M4-M6, S7-S10). The L2 and M6 genes of AHSV serotype 4 (AHSV-4) which encode the outer capsid proteins VP2 and VP5, respectively, were inserted into recombinant baculoviruses downstream of the baculovirus polyhedrin, or p10 promoters. Recombinant baculoviruses expressing VP2, VP5, or VP2 and VP5 proteins of AHSV-4 were isolated. The expressed AHSV proteins were similar in size and antigenic properties to those of viral AHSV-4. Expressed VP2 and VP5 proteins were purified to homogeneity and utilized to differentiate sera from vaccinated and infected horses. Antigens were also used to determine whether any other AHSV serotypes are related to AHSV-4. The results indicated that AHSV-4 is distantly related to some serotypes (e.g., AHSV-2, -6, and -9) but not to others (e.g., AHSV-5 and -7). Hyperimmune monospecific antisera raised in rabbits with purified VP2 neutralized the infectivity of a virulent strain of AHSV-4 isolated from an infected horse during a recent outbreak of the disease in Spain.
Publication Date: 1994-07-01 PubMed ID: 8009847DOI: 10.1006/viro.1994.1351Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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The research article focuses on the role of the outer capsid proteins, VP2 and VP5, of African horsesickness virus (AHSV) in virus neutralization. The study structures the proteins in baculoviruses to use them in differentiating sera from vaccinated and infected horses, as well as determining the relatedness of different AHSV serotypes.
Overview of African horsesickness virus
- AHSV is a virus transmitted through gnats and falls under the Orbivirus genus of the Reoviridae family. Its genome comprises 10 categories of double-stranded RNA species.
Experimentation with AHSV outer capsid proteins
- Using AHSV serotype 4 (AHSV-4), the researchers inserted the genes L2 and M6 – responsible for outer capsid proteins VP2 and VP5, respectively – into recombinant baculoviruses located downstream of baculovirus polyhedrin or p10 promoters.
- They successfully isolated recombinant baculoviruses that expressed VP2, VP5, or both proteins. It was found that these expressed proteins were quite alike in terms of size and antigenic properties to those in viral AHSV-4.
Using expressed proteins for further analysis
- The scientists carried out purification of the expressed VP2 and VP5 proteins and used them to highlight differences between the sera of vaccinated horses and those infected by AHSV.
- The purified antigens were also employed to discover if any other AHSV serotypes bore any relation to AHSV-4.
- The results indicated that AHSV-4 was distantly related to specific serotypes (like AHSV-2, -6, and -9) but not with others (such as AHSV-5 and -7).
Understanding virus neutralization
- Hyperimmune monospecific antisera – made from purified VP2 and administered to rabbits – was found to neutralize the virulence of a particular AHSV-4 strain that was previously isolated from an infected horse during an outbreak in Spain.
- This confirmed the crucial role of VP2 in virus neutralization, validating the main premise of the research study.
Cite This Article
APA
Martinez-Torrecuadrada JL, Iwata H, Venteo A, Casal I, Roy P.
(1994).
Expression and characterization of the two outer capsid proteins of African horsesickness virus: the role of VP2 in virus neutralization.
Virology, 202(1), 348-359.
https://doi.org/10.1006/viro.1994.1351 Publication
Researcher Affiliations
- Institute of Virology and Environmental Microbiology, Oxford, United Kingdom.
MeSH Terms
- African Horse Sickness / immunology
- African Horse Sickness / microbiology
- African Horse Sickness / prevention & control
- African Horse Sickness Virus / classification
- African Horse Sickness Virus / immunology
- African Horse Sickness Virus / physiology
- Animals
- Antibodies, Viral / immunology
- Baculoviridae / genetics
- Base Sequence
- Capsid / immunology
- Capsid / physiology
- Capsid Proteins
- Cell Line
- Cloning, Molecular
- DNA, Viral
- Female
- Horses
- Molecular Sequence Data
- Moths
- Neutralization Tests
- Rabbits
- Recombinant Proteins / chemistry
- Recombinant Proteins / immunology
- Recombinant Proteins / metabolism
- Serotyping
- Solubility
- Transfection
- Vero Cells
Grant Funding
- AI26879 / NIAID NIH HHS
Citations
This article has been cited 10 times.- Jiménez-Cabello L, Utrilla-Trigo S, Barreiro-Piñeiro N, Pose-Boirazian T, Martínez-Costas J, Marín-López A, Ortego J. Nanoparticle- and Microparticle-Based Vaccines against Orbiviruses of Veterinary Importance.. Vaccines (Basel) 2022 Jul 14;10(7).
- Rutkowska DA, Mokoena NB, Tsekoa TL, Dibakwane VS, O'Kennedy MM. Plant-produced chimeric virus-like particles - a new generation vaccine against African horse sickness.. BMC Vet Res 2019 Dec 3;15(1):432.
- Ngoveni HG, van Schalkwyk A, Koekemoer JJO. Evidence of Intragenic Recombination in African Horse Sickness Virus.. Viruses 2019 Jul 18;11(7).
- Aksular M, Calvo-Pinilla E, Marín-López A, Ortego J, Chambers AC, King LA, Castillo-Olivares J. A single dose of African horse sickness virus (AHSV) VP2 based vaccines provides complete clinical protection in a mouse model.. Vaccine 2018 Nov 12;36(46):7003-7010.
- Karamalla ST, Gubran AI, Adam IA, Abdalla TM, Sinada RO, Haroun EM, Aradaib IE. Sero-epidemioloical survey on African horse sickness virus among horses in Khartoum State, Central Sudan.. BMC Vet Res 2018 Aug 1;14(1):230.
- van Gennip RGP, van de Water SGP, Potgieter CA, van Rijn PA. Structural Protein VP2 of African Horse Sickness Virus Is Not Essential for Virus Replication In Vitro.. J Virol 2017 Feb 15;91(4).
- Kanai Y, van Rijn PA, Maris-Veldhuis M, Kaname Y, Athmaram TN, Roy P. Immunogenicity of recombinant VP2 proteins of all nine serotypes of African horse sickness virus.. Vaccine 2014 Sep 3;32(39):4932-7.
- van Wyngaardt W, Mashau C, Wright I, Fehrsen J. Serotype- and serogroup-specific detection of African horsesickness virus using phage displayed chicken scFvs for indirect double antibody sandwich ELISAs.. J Vet Sci 2013;14(1):95-8.
- Martínez-Torrecuadrada JL, Lázaro B, Rodriguez JF, Casal JI. Antigenic properties and diagnostic potential of baculovirus-expressed infectious bursal disease virus proteins VPX and VP3.. Clin Diagn Lab Immunol 2000 Jul;7(4):645-51.
- Martínez-Torrecuadrada JL, Díaz-Laviada M, Roy P, Sánchez C, Vela C, Sánchez-Vizcaíno JM, Casal JI. Serologic markers in early stages of African horse sickness virus infection.. J Clin Microbiol 1997 Feb;35(2):531-5.
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