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Home / Equine Research Bank / Clin Vaccine Immunol / Development and characterization of an infectious cDNA clone...
Clinical and vaccine immunology : CVI2012; 19(8); 1312-1321; doi: 10.1128/CVI.00302-12

Development and characterization of an infectious cDNA clone of the modified live virus vaccine strain of equine arteritis virus.

Abstract: A stable full-length cDNA clone of the modified live virus (MLV) vaccine strain of equine arteritis virus (EAV) was developed. RNA transcripts generated from this plasmid (pEAVrMLV) were infectious upon transfection into mammalian cells, and the resultant recombinant virus (rMLV) had 100% nucleotide identity to the parental MLV vaccine strain of EAV. A single silent nucleotide substitution was introduced into the nucleocapsid gene (pEAVrMLVB), enabling the cloned vaccine virus (rMLVB) to be distinguished from parental MLV vaccine as well as other field and laboratory strains of EAV by using an allelic discrimination real-time reverse transcription (RT)-PCR assay. In vitro studies revealed that the cloned vaccine virus rMLVB and the parental MLV vaccine virus had identical growth kinetics and plaque morphologies in equine endothelial cells. In vivo studies confirmed that the cloned vaccine virus was very safe and induced high titers of neutralizing antibodies against EAV in experimentally immunized horses. When challenged with the heterologous EAV KY84 strain, the rMLVB vaccine virus protected immunized horses in regard to reducing the magnitude and duration of viremia and virus shedding but did not suppress the development of signs of EVA, although these were reduced in clinical severity. The vaccine clone pEAVrMLVB could be further manipulated to improve the vaccine efficacy as well as to develop a marker vaccine for serological differentiation of EAV naturally infected from vaccinated animals.
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Publication Date: 2012-06-27 PubMed ID: 22739697PubMed Central: PMC3416077DOI: 10.1128/CVI.00302-12Google Scholar: Lookup
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Summary

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The research article outlines the development and properties of a stable full-length cDNA clone of a modified live virus (MLV) vaccine strain of the equine arteritis virus (EAV). The research shows the effectiveness of this clone in providing immunity to equine arteritis in horses through producing identical growth kinetics and plaque morphologies in equine endothelial cells as the parental MLV vaccine virus.

Development of the cDNA Clone

  • The research team developed a stable full-length cDNA clone of the MLV vaccine strain of EAV. This plasmid is denoted as pEAVrMLV.
  • They found that RNA transcripts generated from this plasmid were infectious on transfection into mammalian cells.
  • The resultant recombinant virus (rMLV) had a 100% nucleotide identity to the parental MLV vaccine strain of EAV, effectively making it an exact copy of the beneficial properties of the parental vaccine strain.

Introduction of a Single Nucleotide Substitution

  • A silent nucleotide substitution was introduced into the nucleocapsid gene, now referred to as pEAVrMLVB. This modification allowed the cloned vaccine virus (rMLVB) to be distinguished from the parental MLV vaccine and other EAV strains.
  • This differentiation was made possible using an allelic discrimination real-time reverse transcription (RT)-PCR assay, a technique commonly used in gene research for the identification of specific gene sequences.

In Vitro and In Vivo Studies

  • In vitro studies, conducted in a controlled lab setting, revealed the cloned vaccine virus (rMLVB) and the parental MLV vaccine virus had identical growth kinetics and plaque morphologies when tested on equine endothelial cells, effectively mirroring the beneficial attributes the current vaccine provides.
  • In vivo studies, conducted within living organisms, showed that the cloned vaccine virus also safely induced high amounts of neutralizing antibodies against EAV in immunized horses.
  • When the horses were subsequently challenged with a different EAV strain (KY84), the rMLVB vaccine virus effectively protected the immunized horses in reducing the severity of viremia and virus shedding.
  • However, it is important to note that the rMLVB vaccine virus did not fully suppress the development of signs of equine viral arteritis (EVA), though the symptoms were reduced in clinical severity.

Implications and Future Applications

  • The research indicates the potential to further manipulate the vaccine clone pEAVrMLVB to improve vaccine efficacy.
  • This could not only make the vaccine more effective but also aid in the development of a marker vaccine. This would allow for the serological differentiation of EAV naturally infected horses from vaccinated animals, an improvement that would greatly enhance EAV monitoring and control measures.

Cite This Article

APA
Zhang J, Go YY, Huang CM, Meade BJ, Lu Z, Snijder EJ, Timoney PJ, Balasuriya UB. (2012). Development and characterization of an infectious cDNA clone of the modified live virus vaccine strain of equine arteritis virus. Clin Vaccine Immunol, 19(8), 1312-1321. https://doi.org/10.1128/CVI.00302-12

Publication

Clinical and vaccine immunology : CVI
ISSN: 1556-679X
NlmUniqueID: 101252125
Country: United States
Language: English
Volume: 19
Issue: 8
Pages: 1312-1321

Researcher Affiliations

Zhang, Jianqiang
  • Maxwell H. Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Lexington, Kentucky, USA.
Go, Yun Young
    Huang, Chengjin M
      Meade, Barry J
        Lu, Zhengchun
          Snijder, Eric J
            Timoney, Peter J
              Balasuriya, Udeni B R

                MeSH Terms

                • Animals
                • Antibodies, Neutralizing / blood
                • Antibodies, Viral / blood
                • Cells, Cultured
                • DNA, Complementary / genetics
                • Endothelial Cells / virology
                • Equartevirus / classification
                • Equartevirus / genetics
                • Equartevirus / growth & development
                • Genotype
                • Horses
                • Molecular Sequence Data
                • Nucleocapsid / genetics
                • Point Mutation
                • RNA, Viral / genetics
                • Real-Time Polymerase Chain Reaction
                • Reverse Transcriptase Polymerase Chain Reaction
                • Sequence Analysis, DNA
                • Sequence Homology, Nucleic Acid
                • Transcription, Genetic
                • Vaccines, Attenuated / administration & dosage
                • Vaccines, Attenuated / genetics
                • Vaccines, Attenuated / immunology
                • Viral Plaque Assay
                • Viral Vaccines / administration & dosage
                • Viral Vaccines / genetics
                • Viral Vaccines / immunology

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