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

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 disad...

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Main Authors: Shcherbik, Svetlana, Pearce, Nicholas, Kiseleva, Irina, Larionova, Natalie, Rudenko, Larisa, Xu, Xiyan, Wentworth, David E., Bousse, Tatiana
Format: Online
Language:English
Published: 2015
Online Access:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4773654/
id pubmed-4773654
recordtype oai_dc
spelling pubmed-47736542016-03-02 Implementation of new approaches for generating conventional reassortants for live attenuated influenza vaccine based on Russian master donor viruses Shcherbik, Svetlana Pearce, Nicholas Kiseleva, Irina Larionova, Natalie Rudenko, Larisa Xu, Xiyan Wentworth, David E. Bousse, Tatiana Article 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. 2015-10-28 2016-01 /pmc/articles/PMC4773654/ /pubmed/26519883 http://dx.doi.org/10.1016/j.jviromet.2015.10.009 Text en http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
repository_type Open Access Journal
institution_category Foreign Institution
institution US National Center for Biotechnology Information
building NCBI PubMed
collection Online Access
language English
format Online
author Shcherbik, Svetlana
Pearce, Nicholas
Kiseleva, Irina
Larionova, Natalie
Rudenko, Larisa
Xu, Xiyan
Wentworth, David E.
Bousse, Tatiana
spellingShingle Shcherbik, Svetlana
Pearce, Nicholas
Kiseleva, Irina
Larionova, Natalie
Rudenko, Larisa
Xu, Xiyan
Wentworth, David E.
Bousse, Tatiana
Implementation of new approaches for generating conventional reassortants for live attenuated influenza vaccine based on Russian master donor viruses
author_facet Shcherbik, Svetlana
Pearce, Nicholas
Kiseleva, Irina
Larionova, Natalie
Rudenko, Larisa
Xu, Xiyan
Wentworth, David E.
Bousse, Tatiana
author_sort Shcherbik, Svetlana
title Implementation of new approaches for generating conventional reassortants for live attenuated influenza vaccine based on Russian master donor viruses
title_short Implementation of new approaches for generating conventional reassortants for live attenuated influenza vaccine based on Russian master donor viruses
title_full Implementation of new approaches for generating conventional reassortants for live attenuated influenza vaccine based on Russian master donor viruses
title_fullStr Implementation of new approaches for generating conventional reassortants for live attenuated influenza vaccine based on Russian master donor viruses
title_full_unstemmed Implementation of new approaches for generating conventional reassortants for live attenuated influenza vaccine based on Russian master donor viruses
title_sort implementation of new approaches for generating conventional reassortants for live attenuated influenza vaccine based on russian master donor viruses
description 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.
publishDate 2015
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4773654/
_version_ 1613545796491280384

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