Development of a chimeric hemagglutinin-based live-attenuated influenza vaccine against both lineages of influenza B virus

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Abstract

BACKGROUND: The development of a universal influenza vaccine remains a critical goal to enhance protection against diverse influenza virus strains. While vaccines against influenza type A viruses have benefited from the use of chimeric hemagglutinin designs, strategies for influenza type B vaccines still lag.

AIM: The study aimed to investigate the efficacy of chimeric vaccine strains to induce humoral immune response targeting conserved antigenic sites of influenza type B virus.

METHODS: A chimeric hemagglutinin was engineered by combining the head domain from the B/Brisbane/60/2008 virus (Victoria lineage) with the stalk domain from the B/Phuket/3073/2013 virus (Yamagata lineage). The gene encoding the chimeric hemagglutinin was incorporated into a vaccine virus based on the cold-adapted B/USSR/60/69 master donor virus to produce live-attenuated influenza vaccine. Mice were sequentially vaccinated with the conventional live-attenuated influenza vaccine and then with the recombinant live-attenuated influenza vaccine expressing the chimeric hemagglutinin. Immune responses and cross-protection against both homologous and heterologous influenza type B virus strains were assessed.

RESULTS: The engineered chimeric hemagglutinin did not impair the replication or assembly of the vaccine virus. Sequential vaccination induced a robust humoral immune response and provided protection against both homologous and heterologous influenza type B virus strains in the mouse model.

CONCLUSION: Live-attenuated influenza type B vaccines expressing chimeric hemagglutinin show promise in broadening protection against influenza type B virus infection. These findings support the development of a universal influenza type B vaccine using a chimeric hemagglutinin design.

About the authors

Pei-Fong Wong

Institute of Experimental Medicine

Author for correspondence.
Email: po333222@gmail.com
ORCID iD: 0000-0002-7939-6313
SPIN-code: 7151-7480
Russian Federation, Saint Petersburg

Ekaterina A. Stepanova

Institute of Experimental Medicine

Email: fedorova.iem@gmail.com
ORCID iD: 0000-0002-8670-8645
SPIN-code: 8010-3047

Cand. Sci. (Biology)

Russian Federation, Saint Petersburg

Ekaterina A. Bazhenova

Institute of Experimental Medicine

Email: sonya.01.08@mail.ru
ORCID iD: 0000-0003-3280-556X
SPIN-code: 5169-1426

Cand. Sci. (Biology)

Russian Federation, Saint Petersburg

Svetlana A. Donina

Institute of Experimental Medicine

Email: sveta.donina@gmail.com
ORCID iD: 0000-0002-6502-8341
SPIN-code: 6961-3849

Cand. Sci. (Biology)

Russian Federation, Saint Petersburg

Larisa G. Rudenko

Institute of Experimental Medicine

Email: vaccine@mail.ru
ORCID iD: 0000-0002-0107-9959
SPIN-code: 4181-1372

MD, Dr. Sci. (Medicine)

Russian Federation, Saint Petersburg

Irina N. Isakova-Sivak

Institute of Experimental Medicine

Email: isakova.sivak@iemspb.ru
ORCID iD: 0000-0002-2801-1508
SPIN-code: 3469-3600

Dr. Sci. (Biology), Corresponding Member of the RAS

Russian Federation, Saint Petersburg

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Supplementary files

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2. Fig. 1. Design of chimeric HA/B constructs. Influenza B virus HA monomer (based on data reported under PDB accession number 4M44 [22]). The head domain is located between alanines 57 and 305 (B/Yamagata/16/88 numbering, starting with methionine). TM, transmembrane domain; CTD, cytoplasmic tail domain.

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3. Fig. 2. Growth of recombinant LAIV viruses in MDCK cells. Confluent monolayers of MDCK cells were inoculated at an MOI of 0.001 indicated viruses and incubated at 33 °C. Culture supernatants were collected at 0, 24, 48, 72 and 96 hpi and viral titers were quantified by TCID50 assays. Dotted line shows the limit of virus detection in the TCID50 assay.

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4. Fig. 3. Virus replication and tissue tropism of the 6+2 LAIV or BrH viruses in the respiratory tracts of mice. At 3 dpi, four animals from each group were euthanized, and virus titers in the upper respiratory tracts (NT) or lower respiratory tracts (lungs) of the mice were determined by limiting dilutions in eggs.

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5. Fig. 4. Immunogenicity of sequential vaccination with IBV live-attenuated influenza vaccine viruses and/or cHA/BrH virus measured by HI, MN assay, and ELISA against diverse WT IBVs at 43 dpi. Serum responses to Br-wt (a), Ph-wt (b), Lee-wt (c), and Ma-wt (d). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

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6. Fig. 5. Induction of anti-stalk IgG antibodies after sequential vaccination with IBV LAIV viruses and/or cHA/BrH virus. ELISA was performed with a chimeric recombinant H8BY recombinant protein consisting of HA globular head domain from the influenza A/H8N4 virus and HA stalk domain from B/Phuket/3037/2013. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

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7. Fig. 6. Weight loss and survival rate of vaccinated groups challenged with Br-wt (a), Ph-wt (b), Lee-wt (c), and Ma-wt (d) in mice. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

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8. Fig. 7. Protective activity of 6+2 and BrH LAIV viruses in mice. Immunized mice were challenged with B/Brisbane/60/2008 (left panel) and B/Phuket/3037/2013 virus (right panel); viral titers were determined in lungs and nasal turbinates on Day 3 post challenge. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

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