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Influenza Hemagglutinin Head Domain Mimicry by Rational Design

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Abstract

Despite diligent vaccination efforts, influenza virus infection remains a major cause for respiratory-related illness across the globe. The less-than-optimal immunity conferred by the currently prescribed seasonal vaccines and protracted production times warrant the development of novel vaccines. Induction of an epitope-focused antibody response targeting known neutralization epitopes is a viable strategy to enhance the breadth of protection against rapidly evolving infectious viruses. We report the development of a design framework to mimic the hemagglutinin (HA) head fragment of H1-subtype viruses by delineating the interaction network of invariant residues lining the receptor binding site (RBS); a site targeted by cross-reactive neutralizing antibodies. The incorporation of multiple sequence alignment information in our algorithm to fix the construct termini and engineer rational mutations facilitates the facile extension of the design to heterologous (subtype-specific) influenza strains. We evaluated our design protocol by generating head fragments from divergent influenza A H1N1 A/Puerto Rico/8/34 and pH1N1 A/California/07/2009 strains that share a sequence identity of only 74.4% within the HA1 subunit. The designed immunogens exhibited characteristics of a well-ordered protein, and bound conformation-specific RBS targeting antibodies with high affinity, a desirable feature for putative vaccine candidates. Additionally, the bacterial expression of these immunogens provides a low-cost, rapidly scalable alternative.

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Acknowledgements

We thank Dr. James Crowe for providing the 2D1 antibody. The anti-H1N1 A/Puerto Rico/8/34 virus convalescent sera was provided by Dr. Jessica Flynn (Merck Research Laboratories, PA, USA). Dr. Satish Kumar Gupta provided the anti-pH1N1 A/California/07/2009 virus convalescent sera and MA-2077 antibody.

Funding

VVAM acknowledges the fellowship received from the Council of Scientific and Industrial Research, Government of India (GOI). SS is recipient of research associate fellowship from Department of Biotechnology (DBT), GOI. This work was supported in part by a grant from the DBT, GOI to RV.

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  1. V Vamsee Aditya Mallajosyula and Shiv Swaroop have contributed equally to this work.

Authors and Affiliations

  1. Molecular Biophysics Unit, Indian Institute of Science, Bangalore, 560012, India

    V Vamsee Aditya Mallajosyula, Shiv Swaroop & Raghavan Varadarajan

  2. Department of Biochemistry, Central University of Rajasthan, Kishangarh, Ajmer, 305817, India

    Shiv Swaroop

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  1. V Vamsee Aditya Mallajosyula
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  3. Raghavan Varadarajan
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All authors contributed to the study conception and design. Experiments and analysis were performed by V VAM and SS under the supervision of RV. All authors read and approved the final manuscript.

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Correspondence to Raghavan Varadarajan.

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Mallajosyula, V.A., Swaroop, S. & Varadarajan, R. Influenza Hemagglutinin Head Domain Mimicry by Rational Design. Protein J 39, 434–448 (2020). https://doi.org/10.1007/s10930-020-09930-z

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