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Journal of virological methods2009; 163(2); 424-432; doi: 10.1016/j.jviromet.2009.11.006

A multisystem approach for development and evaluation of inactivated vaccines for Venezuelan equine encephalitis virus (VEEV).

Abstract: A multisystem approach was used to assess the efficiency of several methods for inactivation of Venezuelan equine encephalitis virus (VEEV) vaccine candidates. A combination of diverse assays (plaque, in vitro cytopathology and mouse neurovirulence) was used to verify virus inactivation, along with the use of a specific ELISA to measure retention of VEEV envelope glycoprotein epitopes in the development of several inactivated VEEV candidate vaccines derived from an attenuated strain of VEEV (V3526). Incubation of V3526 aliquots at temperatures in excess of 64 degrees C for periods >30 min inactivated the virus, but substantially reduced VEEV specific monoclonal antibody binding of the inactivated material. In contrast, V3526 treated either with formalin at concentrations of 0.1% or 0.5% (v/v) for 4 or 24 h, or irradiated with 50 kGy gamma radiation rendered the virus non-infectious while retaining significant levels of monoclonal antibody binding. Loss of infectivity of both the formalin inactivated (fV3526) and gamma irradiated (gV3526) preparations was confirmed via five successive blind passages on BHK-21 cells. Similarly, loss of neurovirulence for fV3526 and gV3526 was demonstrated via intracerebral inoculation of suckling BALB/c mice. Excellent protection against subcutaneous challenge with VEEV IA/B Trinidad donkey strain was demonstrated using a two dose immunization regimen with either fV3526 or gV3526. The combination of in vitro and in vivo assays provides a practical approach to optimize manufacturing process parameters for development of other inactivated viral vaccines.
2009 Elsevier B.V. All rights reserved.
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Publication Date: 2009-11-10 PubMed ID: 19903494PubMed Central: PMC2815040DOI: 10.1016/j.jviromet.2009.11.006Google Scholar: Lookup
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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 research is about examining multiple methods for inactivating the Venezuelan equine encephalitis virus (VEEV) for use in creating a vaccine. The article discusses the experiment that determines which inactivation method maintains important elements of the virus necessary for effective immunisation.

Research Methodology

The researchers used an integrated approach, applying multiple assays to assess the efficiency of different techniques to inactivate the VEEV for use in vaccines. They utilized:

  • A plaque assay, which helps measure the virus’s ability to infect host cells.
  • An in vitro cytopathology assay to study the morphological changes in host cells due to viral infections.
  • A mouse neurovirulence assay that determined the virus’s potency for creating disease.
  • A specific enzyme-linked immunosorbent assay (ELISA) that helped measure how well vaccines retained VEEV envelope glycoprotein epitopes (the part of the antigen that’s recognized by the immune system).

The various inactivation methods tested included:

  • Heat, where aliquots of V3526, an attenuated strain of VEEV, were incubated at over 64 degrees Celsius for more than 30 minutes.
  • Chemical treatment, where V3526 was exposed to 0.1% or 0.5% formalin for 4 or 24 hours.
  • Radiation, where V3526 was irradiated with a 50 kGy gamma ray dose.

Research Findings

The research found that inactivating the virus with heat led to a significant loss in specific monoclonal antibody binding that is required for a potentially successful vaccine.
In contrast, the formalin-treated or gamma-rays irradiated V3526 remained non-infectious and retained significant antibody binding levels.

The loss of infectivity for formalin-inactivated and gamma-irradiated preparations was confirmed by five successive, blind passages on BHK-21 cells. Similarly, loss of neurovirulence for these preparations was confirmed through intracerebral inoculation of suckling mice.

The research further showed superior protection against a challenge with the VEEV Trinidad donkey strain using a two-dose immunization regimen with either formalin-inactivated or gamma-irradiated V3526.

Research Implications

The research highlights the efficacious use of formaldehyde or gamma irradiation to produce a working vaccine that retains the virus’s necessary attributes. The authors suggest that their multisystem approach to assessing virus inactivation and immunity retention presents an ideal strategy to optimize manufacturing process parameters for the development of other inactivated viral vaccines.

Cite This Article

APA
Fine DL, Jenkins E, Martin SS, Glass P, Parker MD, Grimm B. (2009). A multisystem approach for development and evaluation of inactivated vaccines for Venezuelan equine encephalitis virus (VEEV). J Virol Methods, 163(2), 424-432. https://doi.org/10.1016/j.jviromet.2009.11.006

Publication

Journal of virological methods
ISSN: 1879-0984
NlmUniqueID: 8005839
Country: Netherlands
Language: English
Volume: 163
Issue: 2
Pages: 424-432

Researcher Affiliations

Fine, Donald L
  • DynPort Vaccine Company LLC (DVC), A CSC Company, 64 Thomas Johnson Drive, Frederick, MD 21702, USA. dfine@csc.com
Jenkins, Erin
    Martin, Shannon S
      Glass, Pamela
        Parker, Michael D
          Grimm, Brad

            MeSH Terms

            • Animals
            • Antibodies, Viral / blood
            • Cell Line
            • Cricetinae
            • Disinfectants / pharmacology
            • Encephalitis Virus, Venezuelan Equine / drug effects
            • Encephalitis Virus, Venezuelan Equine / immunology
            • Encephalitis Virus, Venezuelan Equine / pathogenicity
            • Encephalitis Virus, Venezuelan Equine / radiation effects
            • Encephalomyelitis, Venezuelan Equine / virology
            • Enzyme-Linked Immunosorbent Assay / methods
            • Formaldehyde / pharmacology
            • Gamma Rays
            • Hot Temperature
            • Mice
            • Mice, Inbred BALB C
            • Survival Analysis
            • Time Factors
            • Vaccines, Inactivated / immunology
            • Viral Plaque Assay
            • Viral Vaccines / immunology
            • Virulence
            • Virus Cultivation
            • Virus Inactivation

            Grant Funding

            • UC1 AI062538 / NIAID NIH HHS
            • UC1 AI062538-01 / NIAID NIH HHS
            • 1UC1AI062538-01 / NIAID NIH HHS

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