Orthogonal inactivation of influenza and the creation of detergent resistant viral aggregates: towards a novel vaccine strategy

Belanger, Raviv Yossef, Viard Mathias, Baxa Ulrich, Blumenthal, Blumenthal Robert

Description

It has been previously shown that enveloped viruses can be inactivated using aryl azides, such as 1-iodo-5-azidonaphthalene (INA), plus UVA irradiation with preservation of surface epitopes in the inactivated virus preparations. Prolonged UVA irradiation in the presence of INA results in ROS-species formation, which in turn results in detergent resistant viral protein fractions. Results Herein, we characterize the applicability of this technique to inactivate influenza. It is shown that influenza virus + INA (100 micromolar) + UVA irradiation for 30 minutes results in a significant ( p < 0.05) increase in pelletablehemagglutinin after Triton X-100 treatment followed by ultracentrifugation. Additionally, characterization of the virus suspension by immunogold labeling in cryo-EM, and viral pellet characterization via immunoprecipitation with a neutralizing antibody, shows preservation of neutralization epitopes after this treatment. Conclusion These orthogonally inactivated viral preparations with detergent resistant fractions are being explored as a novel route for safe, effective inactivated vaccines generated from a variety of enveloped viruses.

Sujets

• Détergent
• Hemagglutinin
• Tritón
• Vaccine
• Inactivation
• VIH
• Subtypes of HIV
• Orthogonalité

Informations

• Publié par: biomed
• Publié le: 01 janvier 2012
• Nombre de lectures: 23
• Langue: English
• Poids de l'ouvrage: 2 Mo

Extrait

Belanger et al . Virology Journal 2012, 9 :72 http://www.virologyj.com/content/9/1/72

R E S E A R C H Open Access Orthogonal inactivation of influenza and the creation of detergent resistant viral aggregates: towards a novel vaccine strategy Julie M Belanger 1,4 , Yossef Raviv 2 , Mathias Viard 2 , Ulrich Baxa 3 and Robert Blumenthal 1*

Abstract Background: It has been previously shown that enveloped viruses can be inactivated using aryl azides, such as 1- iodo-5-azidonaphthalene (INA), plus UVA irradiation with preservation of surface epitopes in the inactivated virus preparations. Prolonged UVA irradiation in the presence of INA results in ROS-species formation, which in turn results in detergent resistant viral protein fractions. Results: Herein, we characterize the applicability of this technique to inactivate influenza. It is shown that influenza virus + INA (100 micromolar) + UVA irradiation for 30 minutes results in a significant ( p < 0.05) increase in pelletablehemagglutinin after Triton X-100 treatment followed by ultracentrifugation. Additionally, characterization of the virus suspension by immunogold labeling in cryo-EM, and viral pellet characterization via immunoprecipitation with a neutralizing antibody, shows preservation of neutralization epitopes after this treatment. Conclusion: These orthogonally inactivated viral preparations with detergent resistant fractions are being explored as a novel route for safe, effective inactivated vaccines generated from a variety of enveloped viruses. Keywords: Influenza virus, Detergent, Detergent resistance, Hemagglutinin, Triton, Vaccine, Inactivation, Human immunodeficiency virus, HIV-1, Orthogonal

Background on the use of the combination of techniques for safe Universal techniques for th e rapid generation of safe, inactivation. These techniques typically operate on effective vaccines from a variety of viruses are needed to  mechanisms independent of one another, and are con- protect against a host of diseases. Inactivated virus vac- sidered “ orthogonal ” inactivation steps. It is generally cines, which start with infectious material, can be pro- accepted that the additive effect of these steps is used duced through routes such as chemical inactivation. towards the “ 15 logs ” of inactivation suggested for the These types of vaccines are currently being used (such generation of a safe vaccine. as in the USA for Influenza), but continue to have lim-  One typical method for orthogonal inactivation, is the itations, such as the inability to cross-react between generation of “ split virus ” vaccines. These types of vac- viral strains and subtypes. For emerging or ill-character- cines, currently used in some influenza preparations, ized novel pathogens, inactivated vaccines raise the ser- rely on chemical means for the initial inactivation of the ious concern of safety. It is generally agreed that the virus, followed by detergent treatment to “ split ” or solu- required inactivation of virus for use in such vaccines is bilize the virus. This solubilized viral protein preparation at least 15 logs of inactivation [1]. Such a high level of  is then further purified to obtain a specific protein inactivation is difficult to determine, and usually relies (hemagglutinin in the case o f influenza). This purified protein only represents a fraction of the overall native virion, and has been removed from its native environ- * 1 C C e o n r t r e e r sp fo ondence:[e-mail] ment in the viral membrane. r Cancer Research Nanobiology Program, National Cancer Institute Frederick, Frederick, USA Full list of author information is available at the end of the article © 2012 Belanger et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.