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    Design of Immunogens Derived from the Conserved Stem of Influenza Virus Hemagglutinin

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    Bhandari, Nisha
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    Abstract
     
     
    Influenza viruses can cause respiratory illnesses and are very contagious, leading to either pandemics or seasonal epidemics and resulting in significant mortality and morbidity rates. The main symptoms of the influenza virus infection are high fever, cough, sore throat, and rhinitis. In most cases, the illness resolves after two weeks. However, in a person with a weak immune system, it might be fatal. Based on the differences in the matrix protein processing, influenza viruses are classified into four types: A, B, C, and D. Influenza A has a high rate of evolution and causes pandemic outbreaks, which have resulted from viral jumps from the aquatic birds or other animal reservoirs to humans. Influenza B virus has a lower rate of evolution then influenza A, exclusively infects humans, and causes annual epidemics. Influenza C causes only mild infection in humans, and Influenza D is not known to infect humans. Influenza A is further divided into 18 Hemagglutinin (HA) and 9 Neuraminidase (NA) subtypes (Tong et al., 2013). Based on phylogenetic analysis of HA, the Influenza A virus is clustered into two groups: Group 1 and Group 2. Based on the difference in hemagglutinin (HA) surface glycoproteins, Yamagata-like (B/Yamagata/16/88) and Victoria-like (B/Victoria/02/87) are the two lineages of the influenza B virus, co-circulating in the human population since the 1980’s, although B/Yamagata appears to have recently become extinct. H1N1 (Influenza A: Group 1), H3N2(Influenza A: Group 2), and Influenza B (Victoria Lineage) are the currently circulating Influenza virus in humans, while a few incidents of H5N1, H7N2, and H9 infections have also been reported in the past. Previously, in our lab, we designed a headless stem domain immunogens containing the entire stem pf HA from the H3 (H3HA6 (Bommakanti et al, 2010)) and H1 (H1HA6 (Bommakanti et al, 2012) forming inclusion bodies and were aggregation-prone. Later, a smaller soluble stem domain immunogen containing a conserved region of HA2 (H1HA10) was designed (Mallajosyula et al, 2014). This conferred complete homologous protection with partial heterologous protection. The above-designed immunogens did not include all epitope residues of conformation-specific broadly neutralizing antibodies CR9114 and CR6261. To address the shortcomings of our previous constructs, we re-analyzed the HA interaction network and engineered novel immunogens. Here, we report the design of bacterially expressed stem domain immunogens containing all the epitope residues of bnAbs CR9114 and CR6261 and the entire HA2 chain of the Influenza virus. The immunogens were subjected to random mutagenesis library display. Yeast surface display was used in which individual designs (or immunogen libraries) were expressed on the yeast surface and were screened for surface expression and binding to conformationspecific bnAbs. A few clones with improved expression and binding were isolated. These mutants differ from wild-type and have mutations outside of the antibody binding epitopes of both bnAbs CR9114 and CR6261, but showed improved binding to both bnAbs.
     
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    https://etd.iisc.ac.in/handle/2005/6559
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    • Molecular Biophysics Unit (MBU) [302]

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