Kar, U and Khaleeq, S and Garg, P and Bhat, M and Reddy, P and Vignesh, VS and Upadhyaya, A and Das, M and Chakshusmathi, G and Pandey, S and Dutta, S and Varadarajan, R (2022) Comparative Immunogenicity of Bacterially Expressed Soluble Trimers and Nanoparticle Displayed Influenza Hemagglutinin Stem Immunogens. In: Frontiers in Immunology, 13 .
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Abstract
Current influenza vaccines need to be updated annually due to mutations in the globular head of the viral surface protein, hemagglutinin (HA). To address this, vaccine candidates have been designed based on the relatively conserved HA stem domain and have shown protective efficacy in animal models. Oligomerization of the antigens either by fusion to oligomerization motifs or display on self-assembling nanoparticle scaffolds, can induce more potent immune responses compared to the corresponding monomeric antigen due to multivalent engagement of B-cells. Since nanoparticle display can increase manufacturing complexity, and often involves one or more mammalian cell expressed components, it is important to characterize and compare various display and oligomerization scaffolds. Using a structure guided approach, we successfully displayed multiple copies of a previously designed soluble, trimeric influenza stem domain immunogen, pH1HA10, on the ferritin like protein, MsDps2 (12 copies), Ferritin (24 copies) and Encapsulin (180 copies). All proteins were expressed in Escherichia coli. The nanoparticle fusion immunogens were found to be well folded and bound to the influenza stem directed broadly neutralizing antibodies with high affinity. An 8.5 à Cryo-EM map of Msdps2-pH1HA10 confirmed the successful design of the nanoparticle fusion immunogen. Mice immunization studies with the soluble trimeric stem and nanoparticle fusion constructs revealed that all of them were immunogenic, and protected mice against homologous (A/Belgium/145-MA/2009) and heterologous (A/Puerto Rico/8/1934) challenge with 10MLD50 mouse adapted virus. Although nanoparticle display conferred a small but statistically significant improvement in protection relative to the soluble trimer in a homologous challenge, heterologous protection was similar in both nanoparticle-stem immunized and trimeric stem immunized groups. Such rapidly producible, bacterially expressed antigens and nanoparticle scaffolds are useful modalities to tackle future influenza pandemics. Copyright © 2022 Kar, Khaleeq, Garg, Bhat, Reddy, Vignesh, Upadhyaya, Das, Chakshusmathi, Pandey, Dutta and Varadarajan.
| Item Type: | Journal Article |
|---|---|
| Publication: | Frontiers in Immunology |
| Publisher: | Frontiers Media S.A. |
| Additional Information: | The copyright for this article belongs to the Authors. |
| Keywords: | diridavumab; epitope; ferritin; influenza vaccine; Influenza virus hemagglutinin; ketamine; nanoparticle; neutralizing antibody; oligomer; sepivac swe; xylazine; hemagglutinin; influenza vaccine; Influenza virus hemagglutinin, animal cell; animal experiment; animal model; Article; B lymphocyte; binding affinity; circular dichroism; controlled study; cryoelectron microscopy; crystal structure; enzyme linked immunosorbent assay; Escherichia coli; female; immobilized metal affinity chromatography; immune response; immunization; immunogenicity; influenza; MDCK cell line; molecular docking; mouse; Mycobacterium smegmatis; nonhuman; oligomerization; pH; polyacrylamide gel electrophoresis; protein expression; protein purification; size exclusion chromatography; surface plasmon resonance; temperature stress; transmission electron microscopy; vaccination; virus neutralization; virus neutralization test; animal; genetics; human; influenza; mammal, Animals; Ferritins; Hemagglutinin Glycoproteins, Influenza Virus; Hemagglutinins; Humans; Influenza Vaccines; Influenza, Human; Mammals; Mice; Nanoparticles |
| Department/Centre: | Division of Biological Sciences > Molecular Biophysics Unit Others |
| Date Deposited: | 29 Sep 2022 04:52 |
| Last Modified: | 29 Sep 2022 04:52 |
| URI: | https://eprints.iisc.ac.in/id/eprint/76806 |
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