Generation and in vitro characterization of engineered cold-adapted influenza A strains with modified NS gene
- Authors: Chistyakova A.K.1, Prokopenko P.I.2, Krutikova E.V.2, Stepanova E.A.2, Isakova-Sivak I.N.2, Rudenko L.G.2
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Affiliations:
- Peter the Great St. Petersburg Polytechnic University
- Institute of Experimental Medicine
- Issue: Vol 21, No 3 (2021)
- Pages: 153-158
- Section: Conference proceedings
- URL: https://journals.eco-vector.com/MAJ/article/view/77563
- DOI: https://doi.org/10.17816/MAJ77563
- ID: 77563
Cite item
Abstract
BACKGROUND: The high variability of influenza strains and the emergence of new variants of viruses lead to the need for constant updating of the composition of influenza vaccines. One of the options for solving this problem is the development of vaccines with enhanced cross-protection against a wide range of influenza strains. Genetically engineered preparations based on live influenza vaccine can be used for targeted stimulation of the cellular immune response. It has been experimentally established that CTL epitopes inserted into the NS gene of the live influenza vaccine strain cause the activation of lymphocytes and the formation of a pool of resident memory T-cells in the lungs of model animals. It is optimal to use experimentally confirmed immunogenic regions for insertion.
AIM: The aim of this study was to rescue a panel of experimental cold-adapted live attenuated influenza vaccine strains with a modified NS gene using A/Leningrad/134/17/57 backbone and recent influenza strains of H1N1, H3N2 and H7N9 subtypes, and evaluate their properties in vitro.
MATERIALS AND METHODS: A cassette encoding immunogenic, conserved among a wide range of influenza strains T-cell epitopes of the influenza virus PB1 protein restricted by common HLA-allotypes was inserted into the gene encoding the NS1 protein. The modified NS gene was cloned into the pCIPolISapIT influenza virus reverse genetics vector. Chimeric influenza viruses were rescued by transfection of Vero cells by electroporation using a standard 8-plasmid system. The growth characteristics of viruses were assessed in developing chicken embryos.
Results: Three strains were successfully obtained based on the live influenza vaccine master donor virus A/Leningrad/ 134/17/57 with a modified NS gene and influenza viruses of the H1N1, H3N2, H7N9 subtypes. Thus, modification of NS gene by insertion of immunogenic PB1 epitopes did not affect the viability and replicative activity of the rescued chimeric live influenza vaccine strains, regardless of the composition of the surface proteins. The strains replicated well at an optimal temperature, had temperature-sensitive phenotype and were able to grow at low temperature.
CONCLUSIONS: The strains will be further studied as candidates for influenza prophylaxis as an experimental universal influenza vaccine.
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About the authors
Anna K. Chistyakova
Peter the Great St. Petersburg Polytechnic University
Author for correspondence.
Email: anna.k.chistiakova@gmail.com
ORCID iD: 0000-0001-9541-5636
student
Russian Federation, Saint PetersburgPolina I. Prokopenko
Institute of Experimental Medicine
Email: pi.prokopenko@gmail.com
Postgraduate student
Russian Federation, Saint PetersburgElena V. Krutikova
Institute of Experimental Medicine
Email: krutikova.iem@gmal.com
ORCID iD: 0000-0002-0383-2625
SPIN-code: 6330-6128
Cand. Sci. (Biol.), Researcher
Russian Federation, Saint PetersburgEkaterina A. Stepanova
Institute of Experimental Medicine
Email: fedorova.iem@gmail.com
ORCID iD: 0000-0002-8670-8645
SPIN-code: 8010-3047
Cand. Sci. (Biol.), Senior Researcher
Russian Federation, Saint PetersburgIrina N. Isakova-Sivak
Institute of Experimental Medicine
Email: isakova.sivak@iemspb.ru
ORCID iD: 0000-0002-2801-1508
SPIN-code: 3469-3600
Dr. Sci. (Biol.), Head of Laboratory
Russian Federation, Saint PetersburgLarisa G. Rudenko
Institute of Experimental Medicine
Email: vaccine@mail.ru
ORCID iD: 0000-0002-0107-9959
SPIN-code: 4181-1372
Dr. Sci. (Med.), Professor, Honored Scientist of the Russian Federation
Russian Federation, Saint PetersburgReferences
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