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Construction, characterization, and immunization of nanoparticles that display a diverse array of influenza HA trimers

Cohen, Alexander A. and Yang, Zhi and Gnanapragasam, Priyanthi N. P. and Ou, Susan and Dam, Kim-Marie A. and Wang, Haoqing and Bjorkman, Pamela J. (2020) Construction, characterization, and immunization of nanoparticles that display a diverse array of influenza HA trimers. . (Unpublished)

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Current influenza vaccines do not elicit broadly protective immune responses against multiple strains. New strategies to focus the humoral immune response to conserved regions on influenza antigens are therefore required for recognition by broadly neutralizing antibodies. It has been suggested that B cells with receptors that recognize conserved epitopes would be preferentially stimulated through avidity effects by mosaic particles presenting multiple forms of a variable antigen. We adapted SpyCatcher-based platforms, AP205 virus-like particles (VLPs) and mi3 nanoparticles (NPs), to covalently co-display SpyTagged hemagglutinin (HA) trimers from different influenza strains. Here we show successful homotypic and heterotypic conjugation of up to 8 different HA trimers to both VLPs and NPs, and demonstrate that conjugated particles were stable for several weeks of storage. We characterized the HA-VLPs and HA-NPs by cryo-electron tomography to derive the average number of conjugated HAs and their separation distances, and compared immunizations of mosaic and homotypic particles in wild-type mice. Both types of HA particles elicited strong antibody responses, but the mosaic particles did not consistently elicit broader immune responses. We conclude that covalent attachment of HAs from currently-circulating influenza strains represents a viable alternative to current annual influenza vaccine strategies, but in the absence of further modifications, is unlikely to represent a method for making a universal influenza vaccine.

Item Type:Report or Paper (Discussion Paper)
Related URLs:
URLURL TypeDescription Paper
Yang, Zhi0000-0001-8680-3784
Wang, Haoqing0000-0003-0277-3018
Bjorkman, Pamela J.0000-0002-2277-3990
Additional Information:The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license. bioRxiv preprint first posted online Jan. 19, 2020. We thank Mark Howarth (Oxford University) for providing plasmids and advice for VLP expression and purification, Jesse Bloom (Fred Hutchinson) for reagents for the infectious virus neutralization assays, Jost Vielmetter and Pauline Hoffmann at the Caltech Beckman Institute Protein Expression Center for help with protein production, Andrey Malyutin and Songye Chen (Caltech) for help with cryo-EM data collection, Rochelle Diamond and Jamie Tijerina at the Caltech Flow Cytometry/Cell Sorting Facility for help in the flow cytometry experiments and analysis, Jennifer Keeffe for implementation of influenza neutralization assays, Claudia Jette for help with figure preparation, Harry Gristick for VLP and NP images for figures, and Andrew Flyak, Jennifer Keeffe, and Claudia Jette for critical reading of the manuscript. EM was done in the Beckman Institute Resource Center for Transmission Electron Microscopy at Caltech. This work was supported by the National Institute Of Allergy And Infectious Diseases of the National Institutes of Health 1R01AI129784 (PJB) and the National Institutes of Health Grant P50 AI150464 (PJB).
Funding AgencyGrant Number
NIHP50 AI150464
Subject Keywords:Influenza virus, Hemagglutinin nanoparticles, Hemagglutinin VLPs, SpyCatcher, Nanoparticle vaccine, Immunizations, Cryo-electron tomography
Record Number:CaltechAUTHORS:20200121-113054457
Persistent URL:
Official Citation:Construction, characterization, and immunization of nanoparticles that display a diverse array of influenza HA trimers. Alexander A. Cohen, Zhi Yang, Priyanthi Gnanapragasam, Susan Ou, Kim-Marie Dam, Haoqing Wang, Pamela J. Bjorkman. bioRxiv 2020.01.18.911388; doi:
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:100815
Deposited By: Tony Diaz
Deposited On:21 Jan 2020 19:38
Last Modified:16 Sep 2020 22:19

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