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Infection and immunity2001; 69(9); 5709-5715; doi: 10.1128/IAI.69.9.5709-5715.2001

Candidate vaccine against botulinum neurotoxin serotype A derived from a Venezuelan equine encephalitis virus vector system.

Abstract: A candidate vaccine against botulinum neurotoxin serotype A (BoNT/A) was developed by using a Venezuelan equine encephalitis (VEE) virus replicon vector. This vaccine vector is composed of a self-replicating RNA containing all of the VEE nonstructural genes and cis-acting elements and also a heterologous immunogen gene placed downstream of the subgenomic 26S promoter in place of the viral structural genes. In this study, the nontoxic 50-kDa carboxy-terminal fragment (H(C)) of the BoNT/A heavy chain was cloned into the replicon vector (H(C)-replicon). Cotransfection of BHK cells in vitro with the H(C)-replicon and two helper RNA molecules, the latter encoding all of the VEE structural proteins, resulted in the assembly and release of propagation-deficient, H(C) VEE replicon particles (H(C)-VRP). Cells infected with H(C)-VRP efficiently expressed this protein when analyzed by either immunofluorescence or by Western blot. To evaluate the immunogenicity of H(C)-VRP, mice were vaccinated with various doses of H(C)-VRP at different intervals. Mice inoculated subcutaneously with H(C)-VRP were protected from an intraperitoneal challenge of up to 100,000 50% lethal dose units of BoNT/A. Protection correlated directly with serum enzyme-linked immunosorbent assay titers to BoNT/A. The duration of the immunity achieved was tested at 6 months and at 1 year postvaccination, and mice challenged at these times remained refractory to challenge with BoNT/A.
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Publication Date: 2001-08-14 PubMed ID: 11500447PubMed Central: PMC98687DOI: 10.1128/IAI.69.9.5709-5715.2001Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The researchers successfully developed a prototype vaccine against the botulinum neurotoxin serotype A using a Venezuelan equine encephalitis virus vector. The vaccine demonstrated potential in providing immunity against the toxin in mice, with lasting effects observed at 6 months and 1 year after vaccination.

Article Explanation

The researchers focused on developing a vaccine against botulinum neurotoxin serotype A, a potent neurotoxin. For this purpose, they used a vector based on the Venezuelan equine encephalitis (VEE) virus. The modified VEE replicon vector consisted of self-replicating RNA and foreign immunogen genes in place of the virus’ structural genes.

  • The non-toxic end fragment (H(C)) of the botulinum neurotoxin was inserted into the replicon vector. This modified replicon and two other RNA molecules encoding the VEE structural proteins were co-introduced into BHK cells.
  • The cells synthesised and released new particles which carried the modified replicon but were non-propagating. These were known as H(C) VEE replicon particles (H(C)-VRP).
  • Significantly, H(C)-VRPs were able to efficiently produce the desired protein in infected cells, as shown by immunofluorescence and Western Blot analysis.

Vaccine Testing

The effectiveness of the H(C)-VRP was tested in mice which were given varying doses at different time intervals. The primary results showed:

  • Mice given subcutaneous inoculation of H(C)-VRP were protected from a significant dose of botulinum neurotoxin.
  • The level of protection was directly linked to the serum titers obtained via enzyme-linked immunosorbent assays.
  • Re-exposure of animals to the neurotoxin 6 months and 1 year post-vaccination indicated that the vaccine provided long-lasting immunity.

Overall, this study demonstrated the potential of the H(C)-VRP vaccine in offering significant, lasting immunity against botulinum neurotoxin serotype A, confirming its efficacy within the experimental context. The researchers’ innovative approach to vaccine design using a virus replicon vector system represents a promising direction in preventing diseases caused by potent neurotoxins.

Cite This Article

APA
Lee JS, Pushko P, Parker MD, Dertzbaugh MT, Smith LA, Smith JF. (2001). Candidate vaccine against botulinum neurotoxin serotype A derived from a Venezuelan equine encephalitis virus vector system. Infect Immun, 69(9), 5709-5715. https://doi.org/10.1128/IAI.69.9.5709-5715.2001

Publication

Infection and immunity
ISSN: 0019-9567
NlmUniqueID: 0246127
Country: United States
Language: English
Volume: 69
Issue: 9
Pages: 5709-5715

Researcher Affiliations

Lee, J S
  • Virology Division, U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland 21702-5011, USA. John.Lee@det.amedd.army.mil
Pushko, P
    Parker, M D
      Dertzbaugh, M T
        Smith, L A
          Smith, J F

            MeSH Terms

            • Animals
            • Bacterial Vaccines / administration & dosage
            • Bacterial Vaccines / genetics
            • Bacterial Vaccines / immunology
            • Botulinum Toxins, Type A / genetics
            • Botulinum Toxins, Type A / immunology
            • Botulism / prevention & control
            • Cell Line
            • Clostridium botulinum / immunology
            • Clostridium botulinum / metabolism
            • Encephalomyelitis, Venezuelan Equine / genetics
            • Encephalomyelitis, Venezuelan Equine / metabolism
            • Genetic Vectors
            • Mice
            • Mice, Inbred BALB C
            • Replicon / genetics
            • Vaccination
            • Vaccines, Synthetic / administration & dosage
            • Vaccines, Synthetic / immunology

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