IngentaConnect Preparation of genetically engineered AH5N1 and AH7N1 pandemic va...

Authors: Legastelois, Isabelle¹; Garcia-Sastre, Adolfo²; Palese, Peter²; Tumpey, Terrence M.; Maines, Taronna R.³; Katz, Jacqueline M.³; Vogel, Frederick R.¹; Moste, Catherine¹

Source: Influenza and Other Respiratory Viruses, Volume 1, Number 3, May 2007 , pp. 95-104(10)

Publisher: Blackwell Publishing

Abstract:

Background

In case of influenza pandemic, a robust, easy and clean technique to prepare reassortants would be necessary. Objectives

Using reverse genetics, we prepared two vaccine reassortants (A/H5N1 × PR8 and A/H7N1 × PR8) exhibiting the envelope glycoproteins from non-pathogenic avian viruses, A/Turkey/Wisconsin/68 (A/H5N9) and A/Rhea/New Caledonia/39482/93 (A/H7N1) and the internal proteins of the attenuated human virus A/Puerto Rico/8/34 (H1N1). Methods

The transfection was accomplished using a mixture of Vero and chicken embryo cells both of which are currently being used for vaccine manufacturing. Results

This process was reproducible, resulting in consistent recovery of influenza viruses in 6 days. Because it is mainly the A/H5N1 strain that has recently crossed the human barrier, it is the A/PR8 × A/H5N1 reassortant (RG5) that was further amplified, either in embryonated hen eggs or Vero cells, to produce vaccine pre-master seed stocks that met quality control specifications. Safety testing in chickens and ferrets was performed to assess the non-virulence of the reassortant, and finally analysis using chicken and ferret sera immunized with the RG5 virus showed that the vaccine candidate elicited an antibody response cross-reactive with the Hong Kong 1997 and 2003 H5N1 strains but not the Vietnam/2004 viruses. Conclusions

The seeds obtained could be used as part of a pandemic vaccine strain `library' available in case of propagation in humans of a new highly pathogenic avian strain.

Keywords: Influenza; pandemic; reverse genetics; vaccine

Document Type: Research article

DOI: 10.1111/j.1750-2659.2007.00015.x

Affiliations: 1: Research and Development, sanofi pasteur, Marcy L'Etoile, France 2: Department of Microbiology, Mount Sinai School of Medicine (MSSM), New York, NY, USA 3: Influenza Branch, CDC, Atlanta, GA, USA

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Preparation of genetically engineered A/H5N1 and A/H7N1 pandemic vaccine viruses by reverse genetics in a mixture of Vero and chicken embryo cells