Genome composition analysis of reassortant influenza viruses used in seasonal and pandemic live attenuated influenza vaccine

Kiseleva, Irina; Voeten, J.T.M.; Teley, Lisette C. P.; Larionova, Natalie; Дубровина, И. А.; Berdygulova, Zhanna A.; Bazhenova, Ekaterina; Bosch, J. F. van den; Heldens, Jacco; Rudenko, Larisa

doi:10.3103/s0891416811040045

Cited by 11 publications

(4 citation statements)

References 18 publications

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“…The second cloning takes place at 32 °C in the presence of anti-MDV serum. The cloned progeny is then genotyped to find the reassortants containing the 6:2 genome composition ( Desheva Iu et al, 2007 ; Kiseleva et al, 2011a , b ). After a 6:2 reassortant is found, additional rounds of cloning are performed in order to enhance the purity of the reassortant virus progeny.…”

Section: Methodsmentioning

confidence: 99%

Implementation of new approaches for generating conventional reassortants for live attenuated influenza vaccine based on Russian master donor viruses

Shcherbik

Pearce

Kiseleva

et al. 2016

Journal of Virological Methods

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Cold-adapted influenza strains A/Leningrad/134/17/57 (H2N2) and B/USSR/60/69, originally developed in Russia, have been reliable master donors of attenuation for preparing live attenuated influenza vaccines (LAIV). The classical strategy for generating LAIV reassortants is robust, but has some disadvantages. The generation of reassortants requires at least 3 passages under selective conditions after co-infection; each of these selective passages takes six days. Screening the reassortants for a genomic composition traditionally starts after a second limiting dilution cloning procedure, and the number of suitable reassortants is limited. We developed a new approach to shorten process of preparing LAIV seed viruses. Introducing the genotyping of reassortants by pyrosequencing and monitoring sequence integrity of surface antigens starting at the first selective passage allowed specific selection of suitable reassortants for the next cloning procedure and also eliminate one of the group selective passage in vaccine candidate generation. Homogeneity analysis confirmed that reducing the number of selective passages didn’t affect the quality of LAIV seed viruses. Finally, the two-way hemagglutination inhibition test, implemented for all the final seed viruses, confirmed that any amino acid substitutions acquired by reassortants during egg propagation didn’t affect antigenicity of the vaccine. Our new strategy reduces the time required to generate a vaccine and was used to generate seasonal LAIVs candidates for the 2012/2013, 2014/2015, and 2015/2016 seasons more rapidly.

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Section: Methodsmentioning

confidence: 99%

Implementation of new approaches for generating conventional reassortants for live attenuated influenza vaccine based on Russian master donor viruses

Shcherbik

Pearce

Kiseleva

et al. 2016

Journal of Virological Methods

View full text Add to dashboard Cite

show abstract

“…The parental origin of the RNA segments of each reassortant virus was monitored by restriction fragment length polymorphism (RFLP) analysis [ 11 ], or determined by mixed PCR assays [ 12 ]. Nucleotide sequencing was performed using a BigDye Terminator v3.1 cycle sequencing kit and a 3130xl Genetic Analyzer (both from Applied Biosystems, Carlsbad, CA, USA) according to manufacturer’s instructions.…”

Section: Materials and Methodologymentioning

confidence: 99%

Contribution of Neuraminidase of Influenza Viruses to the Sensitivity to Sera Inhibitors and Reassortment Efficiency

et al. 2014

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Live attenuated influenza vaccine (LAIV) represent reassortant viruses with hemagglutinin (HA) and neuraminidase (NA) gene segments inherited from circulating wild-type (WT) parental influenza viruses recommended for inclusion into seasonal vaccine formulation, and the 6 internal protein-encoding gene segments from cold-adapted attenuated master donor viruses (genome composition 6:2). In this study, we describe the obstacles in developing LAIV strains while taking into account the phenotypic peculiarities of WT viruses used for reassortment. Genomic composition analysis of 849 seasonal LAIV reassortants revealed that over 80% of reassortants based on inhibitor-resistant WT viruses inherited WT NA, compared to 26% of LAIV reassortants based on inhibitor-sensitive WT viruses. In addition, the highest percentage of LAIV genotype reassortants was achieved when WT parental viruses were resistant to non-specific serum inhibitors. We demonstrate that NA may play a role in influenza virus sensitivity to non-specific serum inhibitors. Replacing NA of inhibitor-sensitive WT virus with the NA of inhibitor-resistant master donor virus significantly decreased the sensitivity of the resulting reassortant virus to serum heat-stable inhibitors.

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“…The parental origin of the RNA segments of each reassortant virus was monitored by RFLP analysis [16], or determined by mixed PCR assay [17] and partial sequencing.…”

Section: Genome Composition Analysismentioning

confidence: 99%

Live Attenuated Influenza Vaccines against Highly Pathogenic H5N1avian Influenza: Development and Preclinical Characterization

Larionova¹,

Kiseleva²,

Isakova–Sivak³

et al. 2013

J Vaccines Vaccin

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In this paper we describe the development and the outcomes of the preclinical studies of temperature-sensitive and cold-adapted candidates for live attenuated influenza vaccine (LAIV) based on highly pathogenic avian influenza viruses A/H5N1 with pandemic potential. The LAIV candidates were developed by methods of classical reassortment between H2N2 Master Donor Virus for Russian LAIV and H5N1 viruses generated by reverse genetics for inactivated vaccine. These reverse genetically generated viruses were chosen as a source of H5 hemagglutinin and contained a modified protease cleavage site believed to be associated with high virulence. The progeny of reassortment had 7:1 gene composition and were characterized by antigen specificity of the HA of the pandemic virus, a high growth rate in chicken embryos and their parameters of temperature sensitivity and cold adaptation confirmed preserved attenuation of the Master Donor Virus. In addition, one H5N1 LAIV 6:2 reassortant was generated by reverse genetics. When tested in appropriate animal models, all candidates for H5N2 LAIV vaccine were found to be safe, immunogenic and effective in protecting from severe disease, mortality and pathology caused by the homologues lethal challenge virus and they almost completely eliminated challenge virus replication in vaccinated animals. They did not appear to differ in tested properties from the reverse genetically generated H5N1 LAIV 6:2 reassortant which contained wild-type N1-neuraminidase in addition to H5-hemagglutinin. The most promising for further clinical trials has been considered a LAIV candidate to virus A/turkey/Turkey/1/2005 (H5N1, clade 2.2).

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Genome composition analysis of reassortant influenza viruses used in seasonal and pandemic live attenuated influenza vaccine

Cited by 11 publications

References 18 publications

Implementation of new approaches for generating conventional reassortants for live attenuated influenza vaccine based on Russian master donor viruses

Implementation of new approaches for generating conventional reassortants for live attenuated influenza vaccine based on Russian master donor viruses

Contribution of Neuraminidase of Influenza Viruses to the Sensitivity to Sera Inhibitors and Reassortment Efficiency

Live Attenuated Influenza Vaccines against Highly Pathogenic H5N1avian Influenza: Development and Preclinical Characterization

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