Expression of the major core antigen VP7 of African horsesickness virus by a recombinant baculovirus and its use as a group-specific diagnostic reagent.
Abstract: The major core protein, VP7, of African horsesickness virus serotype 4 (AHSV-4), the aetiological agent of a recent outbreak of the disease in southern Europe, was expressed in insect cells infected with a recombinant baculovirus containing a cloned copy of the relevant AHSV gene (S7). Analyses of its biochemical and antigenic properties confirmed the authenticity of the protein expressed. The high-level expression of VP7 under the control of the strong polyhedrin promoter of Autographa californica nuclear polyhedrosis virus induced disc-shaped crystals in infected insect cells. This enabled us to purify the protein by a one-step ultracentrifugation procedure and to utilize it for the detection of antibodies raised in horses to various serotypes of AHSV. A serological relationship between AHSV and two other orbiviruses, bluetongue virus and epizootic haemorrhagic disease virus, was also demonstrated.
Publication Date: 1992-04-01 PubMed ID: 1378881DOI: 10.1099/0022-1317-73-4-925Google 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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This research presents the successful production of the chief protein (VP7) of the African horsesickness virus in insect cells and how this protein can be used to detect antibodies in horses, serving as a diagnostic tool for the disease.
Expression of VP7 Protein in Insect Cells
- The research successfully manipulated a recombinant baculovirus, or an artificial virus constructed in the lab, to express the VP7 protein found in the African horsesickness virus (AHSV).
- The AHSV type used in this study was serotype 4 (AHSV-4), which had recently caused an outbreak in Southern Europe.
- The artificial virus carried a cloned copy of the AHSV gene responsible for VP7 production.
Attributes of the Expressed Protein
- Both biochemical and antigenic testing confirmed the expressed VP7 protein’s authenticity, matching the one naturally produced by AHSV.
- The expression of VP7 protein was regulated by the strong polyhedrin promoter of another virus, Autographa californica nuclear polyhedrosis virus.
- The VP7 protein’s production led to the formation of disc-shaped crystals within the infected insect cells.
Purification, Utilization and Serological Relationship
- The researchers were able to purify the VP7 protein using a one-step ultracentrifugation procedure, simplifying the process.
- The purified VP7 protein was then used as a diagnostic reagent to detect antibodies raised by various AHSV serotypes in horses, indicating infection.
- The study also showed a serological relationship, or antibody resemblance, between AHSV and two other orbiviruses – the bluetongue virus and epizootic haemorrhagic disease virus. This indicates these viruses may interact with the immune system in a similar manner.
Cite This Article
APA
Chuma T, Le Blois H, Sánchez-Vizcaíno JM, Diaz-Laviada M, Roy P.
(1992).
Expression of the major core antigen VP7 of African horsesickness virus by a recombinant baculovirus and its use as a group-specific diagnostic reagent.
J Gen Virol, 73 ( Pt 4), 925-931.
https://doi.org/10.1099/0022-1317-73-4-925 Publication
Researcher Affiliations
- University of Alabama, Birmingham 35294.
MeSH Terms
- African Horse Sickness / diagnosis
- African Horse Sickness Virus / genetics
- African Horse Sickness Virus / immunology
- Animals
- Antigens, Viral / biosynthesis
- Antigens, Viral / immunology
- Baculoviridae / genetics
- Base Sequence
- Cells, Cultured
- Cross Reactions
- Epitopes / immunology
- Genetic Vectors
- Molecular Sequence Data
- Recombinant Proteins / biosynthesis
- Recombination, Genetic
- Reoviridae / immunology
- Serotyping
- Viral Core Proteins / biosynthesis
- Viral Core Proteins / genetics
- Viral Core Proteins / immunology
Grant Funding
- A126879 / PHS HHS
Citations
This article has been cited 13 times.- Bekker S, Potgieter CA, van Staden V, Theron J. Investigating the Role of African Horse Sickness Virus VP7 Protein Crystalline Particles on Virus Replication and Release.. Viruses 2022 Oct 4;14(10).
- Bekker S, Huismans H, van Staden V. Generation of a Soluble African Horse Sickness Virus VP7 Protein Capable of Forming Core-like Particles.. Viruses 2022 Jul 26;14(8).
- Dennis SJ, Meyers AE, Hitzeroth II, Rybicki EP. African Horse Sickness: A Review of Current Understanding and Vaccine Development.. Viruses 2019 Sep 11;11(9).
- Dennis SJ, O'Kennedy MM, Rutkowska D, Tsekoa T, Lourens CW, Hitzeroth II, Meyers AE, Rybicki EP. Safety and immunogenicity of plant-produced African horse sickness virus-like particles in horses.. Vet Res 2018 Oct 11;49(1):105.
- Dennis SJ, Meyers AE, Guthrie AJ, Hitzeroth II, Rybicki EP. Immunogenicity of plant-produced African horse sickness virus-like particles: implications for a novel vaccine.. Plant Biotechnol J 2018 Feb;16(2):442-450.
- Fowler VL, Howson ELA, Flannery J, Romito M, Lubisi A, Agüero M, Mertens P, Batten CA, Warren HR, Castillo-Olivares J. Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Assay for the Rapid Detection of African Horse Sickness Virus.. Transbound Emerg Dis 2017 Oct;64(5):1579-1588.
- 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.
- Li JK. Oncolytic bluetongue viruses: promise, progress, and perspectives.. Front Microbiol 2011;2:46.
- Tan BH, Nason E, Staeuber N, Jiang W, Monastryrskaya K, Roy P. RGD tripeptide of bluetongue virus VP7 protein is responsible for core attachment to Culicoides cells.. J Virol 2001 Apr;75(8):3937-47.
- 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.
- Kimman TG, de Leeuw O, Kochan G, Szewczyk B, van Rooij E, Jacobs L, Kramps JA, Peeters B. An indirect double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) using baculovirus-expressed antigen for the detection of antibodies to glycoprotein E of pseudorabies virus and comparison of the method with blocking ELISAs.. Clin Diagn Lab Immunol 1996 Mar;3(2):167-74.
- Basak AK, Gouet P, Grimes J, Roy P, Stuart D. Crystal structure of the top domain of African horse sickness virus VP7: comparisons with bluetongue virus VP7.. J Virol 1996 Jun;70(6):3797-806.
- Cloete M, du Plessis DH, van Dijk AA, Huismans H, Viljoen GJ. Vaccinia virus expression of the VP7 protein of South African bluetongue virus serotype 4 and its use as an antigen in a capture ELISA.. Arch Virol 1994;135(3-4):405-18.
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