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

Virology Journal

Robert Blumenthal³  Ulrich Baxa¹  Mathias Viard⁴  Yossef Raviv⁴  Julie M Belanger² 

[1] Advanced Technology Program, SAIC-Frederick, Inc., NCI-Frederick, Frederick, Maryland 21702, USA;Department of Chemistry and Physics, King's College, Wilkes-Barre, PA, USA;Center for Cancer Research Nanobiology Program, National Cancer Institute Frederick, Frederick, USA;Basic Research Program, SAIC-Frederick, Inc., NCI-Frederick, Frederick, Maryland 21702, USA

关键词: Orthogonal;    HIV-1;    Human immunodeficiency virus;    Inactivation;    Vaccine;    Triton;    Hemagglutinin;    Detergent resistance;    Detergent;    Influenza virus;   

Others  :  1154834
DOI  :  10.1186/1743-422X-9-72

 received in 2011-10-17, accepted in 2012-03-26,  发布年份 2012

【 摘 要 】

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.

【 授权许可】

   
2012 Belanger et al; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Schultz AM, Koff CW, Lawrence DN: Conference reports: workshop on HIV inactivated vaccines. Vaccine 1990, 8:516-517.
• [2]Geeraedts F, Bungener L, Pool J, ter Veer W, Wilschut J, Huckriede A: Whole inactivated virus influenza vaccine is superior to subunit vaccine in inducing immune responses and secretion of proinflammatory cytokines by DCs. Influenza Other Respi Viruses 2008, 2:41-51.
• [3]KarlssonHedestam GB, Fouchier RAM, Phogat S, Burton DR, Sodroski J, Wyatt RT: The challenges of elliciting neutralizing antibodies to HIV-1 and to influenza virus. Nat Rev Microbiol 2008, 6:143-155.
• [4]Bercovici T, Gitler C: 5-[125I] Iodonaphthylazide, a reagent to determine the penetration of proteins into the lipid bilayer of biological membranes. Biochemistry 1978, 17:1484-1489.
• [5]Warfield KL, Swenson DL, Olinger GG, Kalina WV, Viard M, Aitichou M, Chi X, Ibrahim S, Blumenthal R, Raviv Y, et al.: Ebola virus inactivation with preservation of antigenic and structural integrity by a photoinducible alkylating agent. J Infect Dis 2007, 196:S276-S283.
• [6]Raviv Y, Blumenthal R, Tompkins SM, Humberd J, Hogan RJ, Viard M: Hydrophobic inactivation of influenza viruses confers preservation of viral structure with enhanced immunogenicity. J Virol 2008, 82:4612-4619.
• [7]Raviv Y, Viard M, Bess JW Jr, Chertova E, Blumenthal R: Inactivation of retroviruses with preservation of structural integrity by targeting the hydrophobic domain of the viral envelope. J Virol 2005, 79:12394-12400.
• [8]Belanger JM, Raviv Y, Viard M, de la Cruz JM, Nagashima K, Blumenthal R: Characterization of the effects of aryl-azido compounds and UVA irradiation on the viral proteins and infectivity of human immunodeficiency virus type 1. PhotochemPhotobiol 2010, 86:1099-1108.
• [9]Sharma A, Raviv Y, Puri A, Viard M, Blumenthal R, Maheshwari RK: Complete inactivation of venezuelan equine encephalitis virus by 1,5-iodonaphthylazide. BiochemBiophys Res Commun 2007, 358:392-398.
• [10]Belanger JM, Raviv Y, Viard M, de la Cruz JM, Nagashima K, Blumenthal R: Effects of UVA irradiation, aryl azides, and reactive oxygen species on the orthogonal inactivation of the human immunodeficiency virus (HIV-1). Virology 2011, 417(1):221-228.
• [11]Welling GW, van der Zee R, Welling-Wester S: HPLC of Membrane Proteins. In Chromatographic Science Series. Volume Volume 87. secondth edition. Edited by Gooding KM, Regnier FE. New York: Marcel Dekker, Inc; 2002::500-530.
• [12]Yu J, Fischman DA, Steck TL: Selective solubilization of proteins and phospholipids from red blood cell membranes by nonionic detergents. J Supramol Structure 1973, 1:233-248.
• [13]Kirkpatrick FH, Gordesky SE, Marinetti GV: Differential solubilization of proteins, phospholipids and cholesterol of erythrocyte membranes by detergents. BiochimBiophysActa 1974, 345:154-161.
• [14]Carrasco M, Amorium MJ, Digard P: Lipid raft-dependent targeting of the influenza A virus nucleoprotein to the apical plasma membrane. Traffic 2004, 5:979-992.
• [15]Wraith DC, Askonas BA: Induction of influenza A virus cross-reactive cytotoxic t cells by a nuceloprotein/haemagglutinin preparation. J Gen Virol 1985, 66:1327-1331.
• [16]Jonges M, Liu WM, van der Vries E, Jacobi R, Pronk I, Boog C, Koopmans M, Meijer A, Soethout E: Influenza virus inactivation for studies of antigenicity and phenotypic neuraminidase inhibitor resistance profiling. J ClinMicrobiol 2010, 48:928-940.
• [17]Lutz A, Dyall J, Olivo PD, Pekosz A: Virus-inducible reporter genes as a tool for detecting and quantifying influenza A virus replication. J Virol Methods 2005, 126:13-20.
• [18]Product information sheet for NR-3148 BEI Resources
• [19]Szretter KJ, Balish AL, Katz JM: UNIT 15G.1: Influenza: Propagation, Quantification, and Storage. CurrProtMicrobiol 2006, 15G.11.11-15G.11.22.
• [20]Matrosovich M, Matrosovich T, Garten W, Klenk H-D: New low-viscosity overlay medium for viral plaque assays. Virology Journal 2006., 3