Influenza viruses can cause severe respiratory infections in humans, leading to nearly half a million deaths worldwide each year. Improved antiviral drugs are needed to address the threat of development of novel pandemic strains. Current therapeutic interventions target three key proteins in the viral life cycle: neuraminidase, the M2 channel and RNA-dependent-RNA polymerase. Protein-protein interactions between influenza polymerase subunits are potential new targets for drug development. Using a newly developed assay based on AlphaScreen technology, we screened a peptide panel for protein-protein interaction inhibitors to identify a minimal PB1 subunit-derived peptide that retains high inhibition potential and can be further modified. Here, we present an X-ray structure of the resulting decapeptide bound to the C-terminal domain of PA polymerase subunit from pandemic isolate A/California/07/2009 H1N1 at 1.6 Å resolution and discuss its implications for the design of specific, potent influenza polymerase inhibitors.

• MeSH
• antivirové látky farmakologie   MeSH
• interakční proteinové domény a motivy účinky léků   fyziologie   MeSH
• krystalizace MeSH
• lidé MeSH
• RNA-dependentní RNA-polymerasa chemie   metabolismus   MeSH
• vazba proteinů MeSH
• virové proteiny antagonisté a inhibitory   chemie   metabolismus   MeSH
• virus chřipky A, podtyp H1N1 účinky léků   enzymologie   metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND/AIMS: The immune response to influenza vaccine may be influenced by many factors, e.g. age, comorbidities or inflammation, and iron status. METHODS: We studied the vaccine-induced production of hemagglutination-inhibition antibodies (HI) in 133 hemodialysis patients (HD) and 40 controls. To identify variables associated with the immune response, uni- and multivariate regression analyses were performed with seroconversion in HI titers as a dependent variable, with demographics, comorbidities, previous vaccination, inflammation, and iron status as independent variables. RESULTS: Seroconversion rates were lower in HD than in controls [43% versus 73% (H1N1 strain; p < 0.05); 43% versus 53% (H3N2; P=NS); 36% versus 62% (B; p < 0.05)]. In both HD and control groups, the predictors of the inferior HI production were pre-vaccination seroprotection, vaccination in the previous season, and old age. We did not find associations between seroconversion rates and inflammation and iron status in the studied populations. This was also true for a subanalysis of patients without pre-vaccination seroprotection. CONCLUSION: The influenza vaccine-induced antibody production was lower in HD than in controls and was independent of inflammation and iron status in both groups. Besides dependence on dialysis, the variables associated with inferior seroconversion rates included pre-vaccination seroprotection, previous vaccination, and old age.

• MeSH
• buněčná imunita účinky léků   imunologie   MeSH
• dialýza ledvin trendy   MeSH
• lidé středního věku MeSH
• lidé MeSH
• prediktivní hodnota testů MeSH
• senioři MeSH
• vakcinace trendy   MeSH
• vakcíny proti chřipce aplikace a dávkování   MeSH
• věkové faktory MeSH
• virus chřipky A, podtyp H1N1 imunologie   metabolismus   MeSH
• virus chřipky A, podtyp H3N2 imunologie   metabolismus   MeSH
• zánět krev   diagnóza   imunologie   MeSH
• železo krev   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky kontrolované MeSH
• multicentrická studie MeSH
• práce podpořená grantem MeSH

The global outbreak of novel A/H1N1 spread in human population worldwide has revealed an emergency need for producing a vaccine against this virus. Current influenza vaccines encounter problems with safety issues and weak response in high-risk population. It has been established that haemagglutinin is the most important viral antigen to which antibody responses are directed, and recombinant subunit vaccines, haemagglutinin of influenza A and B viruses, have been considered in order to facilitate vaccine production. In the present study, we have focused on construction of a recombinant baculovirus encoding the large subunit of novel influenza virus A/H1N1 haemagglutinin. The full genome of haemagglutinin was cloned into pGEM-TEasy vector and sequenced. The large subunit of the haemagglutinin gene was amplified by PCR using specific primers and cloned into pFast- BacHTc donor plasmid, which was then confirmed by restriction enzyme analysis and sequencing and transformed into E. coli DH10Bac competent cells. The bacmid DNA was transfected into insect cells to produce recombinant baculovirus. Expression of recombinant haemagglutinin in insect cells was determined by SDS-PAGE and immunoblotting. It has been shown that the recombinant haemagglutinin (rHA) obtained from the baculovirus insect cell expression system has suitable immunogenicity in human and can be considered as a candidate flu vaccine. Here we produced large amounts of the HA1 protein of novel influenza A/H1N1 (Iranian isolate) in insect cells. The immunogenicity and efficacy of the recombinant HA1 will be evaluated as a vaccine candidate and compared to the recombinant HA1 produced in a prokaryotic system.

• MeSH
• Baculoviridae genetika   metabolismus   MeSH
• buněčné linie MeSH
• epitopy metabolismus   MeSH
• hemaglutininové glykoproteiny viru chřipky genetika   imunologie   metabolismus   MeSH
• imunoblotting MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• rekombinantní proteiny genetika   metabolismus   MeSH
• sekvenční analýza DNA MeSH
• Spodoptera genetika   virologie   MeSH
• transfekce MeSH
• virus chřipky A, podtyp H1N1 genetika   imunologie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH