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Saint Petersburg, Russia

Mirgorodskaya O.A.,Research Institute of Influenza | Korner R.,Max Planck Institute of Biochemistry | Kozmin Y.P.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Roepstorff P.,University of Southern Denmark
Methods in Molecular Biology | Year: 2012

Amino acid analysis is among the most accurate methods for absolute quantification of proteins and peptides. Here, we combine acid hydrolysis with the addition of isotopically labeled standard amino acids and analysis by mass spectrometry for accurate and sensitive protein quantitation. Quantitation of less than 10 fmol of protein standards with errors below 10% has been demonstrated using this method (1). Source

Koel B.F.,Erasmus University Rotterdam | Burke D.F.,University of Cambridge | Bestebroer T.M.,Erasmus University Rotterdam | Van Der Vliet S.,Erasmus University Rotterdam | And 17 more authors.
Science | Year: 2013

The molecular basis of antigenic drift was determined for the hemagglutinin (HA) of human influenza A/H3N2 virus. From 1968 to 2003, antigenic change was caused mainly by single amino acid substitutions, which occurred at only seven positions in HA immediately adjacent to the receptor binding site. Most of these substitutions were involved in antigenic change more than once. Equivalent positions were responsible for the recent antigenic changes of influenza B and A/H1N1 viruses. Substitution of a single amino acid at one of these positions substantially changed the virus-specific antibody response in infected ferrets. These findings have potentially far-reaching consequences for understanding the evolutionary mechanisms that govern influenza viruses. Source

Plotnikova M.A.,Research Institute of Influenza | Klotchenko S.A.,Research Institute of Influenza | Vasin A.V.,Polytechnic University of Mozambique
Journal of Immunological Methods | Year: 2016

Cytokines are global mediators of cellular communications that are involved in broad array of biological processes, including the immunological and inflammatory mechanisms of virus-host interactions. Measuring the gene expression of simultaneously expressed cytokines is necessary for understanding the pathogenesis of many viral infections, including influenza. We developed a multiplex quantitative real-time PCR (qPCR) method for the detection of the following human cytokines: IL-1B, IL-2, IL-4, IL-6, IL-10, IL-12B, IL-18, IFN-γ and TNF. The assay consisted of three sets of multiple qPCRs; in each qPCR, three target cytokines and reference GAPDH genes were amplified. The assay provided a precise and sensitive quantification of cytokine gene expression with a 20 fmol limit of detection and a 1.5% coefficient of variation. This method was successfully applied to cytokine profiling in epithelial A549 cells that were infected with A/California/07/09 (H1N1pdm2009) virus. © 2016 Elsevier B.V.. Source

Sergeeva M.V.,Research Institute of Influenza | Romanova J.R.,Research Institute of Influenza
Voprosy Virusologii | Year: 2013

Worldwide spreading of H5 and H7 highly pathogenic influenza viruses of the avian origin, which periodically Infect and kill humans without prior adaptation, poses a constant threat of the new pandemic. The effectiveness of the pandemic prevention completely depends on the quality of the existing Influenza vaccines. Typical methods of the vaccine production from the antigenically relevant strains are problematic in case of high virulent H5 and H7 viruses. Therefore, new approaches to the construction of the vaccine strains and production technologies are required In order to protect the population. Source

Rudenko L.,Institute of Experimental Medicine | Isakova-Sivak I.,Institute of Experimental Medicine | Naykhin A.,Institute of Experimental Medicine | Kiseleva I.,Institute of Experimental Medicine | And 6 more authors.
The Lancet Infectious Diseases | Year: 2016

Background: H7N9 avian influenza viruses characterised by high virulence and presence of mammalian adaptation markers have pandemic potential. Specific influenza vaccines remain the main defence. We assessed the safety and immunogenicity of an H7N9 live attenuated influenza vaccine (LAIV) candidate in healthy adult volunteers. Methods: We did a phase 1, double-blind, randomised, placebo-controlled trial in Saint Petersburg, Russia. Eligible participants were healthy adults aged 18-49 years. The participants were randomised 3:1 to receive live vaccine or placebo, according to a computer-generated randomisation scheme. Two doses of vaccine or placebo were administered intranasally 28 days apart, each followed by 7 day stays in hospital. Immune responses were assessed in nasal swabs, saliva, and serum specimens collected before and 28 days after each vaccine dose. The primary outcome was the safety profile. This trial is registered with ClinicalTrials.gov, number NCT02480101. Findings: Between Oct 21, 2014, and Oct 31, 2014, 40 adults were randomised, of whom 39 (98%) were included in the per-protocol analysis (29 in the vaccine group and ten in the placebo group). The frequency of adverse events did not differ between the vaccine and placebo groups. Seroconversion of neutralising antibodies was seen in 14 participants after the first vaccine dose (48%, 95% CI 29·4-67·5) and 21 after the second vaccine dose (72%, 52·8-87·3). Immune responses were seen in 27 of 29 recipients (93%, 95% CI 77·2-99·2). Adverse effects were seen in 19 (63%) vaccine recipients and nine (90%) placebo recipients after the first dose and in nine (31%) and four (40%), respectively, after the second dose. These effects were mainly local and all were mild. Interpretation: The H7N9 LAIV was well tolerated and safe and showed good immunogenicity. Funding: WHO. © 2016 World Health Organization. Source

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