Inaccuracies in prediction of circulating viral strain genotypes and the possibility of novel reassortants causing a pandemic outbreak necessitate the development of an anti-influenza vaccine with increased breadth of protection and potential for rapid production and deployment. The hemagglutinin (HA) stem is a promising target for universal influenza vaccine as stem-specific antibodies have the potential to be broadly cross-reactive towards different HA subtypes. Here, we report the design of a bacterially expressed polypeptide that mimics a H5 HA stem by protein minimization to focus the antibody response towards the HA stem. The HA mini-stem folds as a trimer mimicking the HA prefusion conformation. It is resistant to thermal/chemical stress, and it binds to conformation-specific, HA stem-directed broadly neutralizing antibodies with high affinity. Mice vaccinated with the group 1 HA mini-stems are protected from morbidity and mortality against lethal challenge by both group 1 (H5 and H1) and group 2 (H3) influenza viruses, the first report of cross-group protection. Passive transfer of immune serum demonstrates the protection is mediated by stem-specific antibodies. Furthermore, antibodies indudced by these HA stems have broad HA reactivity, yet they do not have antibody-dependent enhancement activity

A Schneemann

AH Ellebedy

C Ganesh

CJ Wei

D Corti

D Sharma

DC Ekiert

DG Higgins

DJ DiLillo

DT O’Hagan

F Ferrara

F Krammer

G Bommakanti

J Stevens

J Sui

JJ Skehel

JR Whittle

MB Mazanec

NJ Temperton

PH Goff

RD de Vries

S Guthe

S Jegaskanda

S Khurana

SA Valkenburg

TJ Wohlbold

U Rathore

VV Mallajosyula

W Gerhard

Y Huang

Y Wu

Scientific Reports

English

PubMed

Valkenburg, Sophie A

Mallajosyula, Vamsee Aditya V

Li, Olive TW

Chin, Alex WH

Carnell, George

Temperton, Nigel

Varadarajan, Raghavan

Poon, Leo LM

Open Access Repository of IISc Research Publications

Stalking influenza by vaccination with pre-fusion headless HA mini-stem

Mallajosyula, V Vamsee Aditya

Li, Olive T W

Chin, Alex W H

Temperton, Nigel J.

Poon, Leo L M

Kent Academic Repository

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Stalking influenza by vaccination with pre-fusion headless HA mini-stem.

Open Access ArticleAntigenic drift of the HA head necessitates seasonal influenza vaccination, therefore in a bid to develop a universal vaccine, we adopted the approach of protein minimization of the HA protein to focus the antibody response towards the functionally conserved epitopes of the HA stem, using a H5 and H1 based vaccine in a mouse model. The H5 HA mini-stem protein refolds as a trimer, in a highly stable pre-fusion conformation representing the native conformation of the HA stem. The HA mini-stem is biochemically stable under diverse stress conditions, rendering the vaccine compatible for bulk production from E. Coli and pandemic stockpiling. Due to the ideal conformational nature of our HA mini-stem, antibodies elicited by the vaccine competed with other broadly neutralizing stem-specific antibodies (CR6261, F10 and FI6v3). Vaccine stem-specific antibodies were broadly reactive against HA proteins, binding with high affinity. Ultimately, vaccination of mice with the H1 and H5 HA mini-stem resulted in significant protection from morbidity and mortality against challenge with homologous and heterologous influenza viruses, including group 2 influenza, H3N2 challenge. Ours study is the first report of a Group 1 HA-stem vaccine showing Group 2 protection. Passive immune sera transfer demonstrated protection was mediated by vaccine stem-specific antibodies. Virus micro-neutralization was observed by vaccine immune sera for H1N1, H7N7 and H3N2 viruses, but not H5N1 viruses, despite clear protection in vaccinated and passive transfer experiments for H5N1 viruses. This raises important issues for the role of stem-specific antibodies, and whether neutralization is a requisite for board spectrum protection. We believe that this observation will provide critical research directions for developing novel universal influenza vaccines.link_to_OA_fulltex

Mallajosyula, VA

Temperton, N

Carnell, G

Li, OTW

Poon, LML

Valkenburg, SA

Varadarajan, R

Chin, WH

HKU Scholars Hub

Inaccuracies in prediction of circulating viral strain genotypes and the possibility of novel reassortants causing a pandemic outbreak necessitate the development of an anti-influenza vaccine with increased breadth of protection and potential for rapid production and deployment. The hemagglutinin (HA) stem is a promising target for universal influenza vaccine as stem-specific antibodies have the potential to be broadly cross-reactive towards different HA subtypes. Here, we report the design of a bacterially expressed polypeptide that mimics a H5 HA stem by protein minimization to focus the antibody response towards the HA stem. The HA mini-stem folds as a trimer mimicking the HA prefusion conformation. It is resistant to thermal/chemical stress, and it binds to conformation-specific, HA stem-directed broadly neutralizing antibodies with high affinity. Mice vaccinated with the group 1 HA mini-stems are protected from morbidity and mortality against lethal challenge by both group 1 (H5 and H1) and group 2 (H3) influenza viruses, the first report of cross-group protection. Passive transfer of immune serum demonstrates the protection is mediated by stem-specific antibodies. Furthermore, antibodies induced by these HA stems have broad HA reactivity, yet they do not have antibody-dependent enhancement activity.published_or_final_versio

Doak, SA

Mallajosyula, VV

AbstractInaccuracies in prediction of circulating viral strain genotypes and the possibility of novel reassortants causing a pandemic outbreak necessitate the development of an anti-influenza vaccine with increased breadth of protection and potential for rapid production and deployment. The hemagglutinin (HA) stem is a promising target for universal influenza vaccine as stem-specific antibodies have the potential to be broadly cross-reactive towards different HA subtypes. Here, we report the design of a bacterially expressed polypeptide that mimics a H5 HA stem by protein minimization to focus the antibody response towards the HA stem. The HA mini-stem folds as a trimer mimicking the HA prefusion conformation. It is resistant to thermal/chemical stress and it binds to conformation-specific, HA stem-directed broadly neutralizing antibodies with high affinity. Mice vaccinated with the group 1 HA mini-stems are protected from morbidity and mortality against lethal challenge by both group 1 (H5 and H1) and group 2 (H3) influenza viruses, the first report of cross-group protection. Passive transfer of immune serum demonstrates the protection is mediated by stem-specific antibodies. Furthermore, antibodies indudced by these HA stems have broad HA reactivity, yet they do not have antibody-dependent enhancement activity.</jats:p

Sophie A. Valkenburg

V. Vamsee Aditya Mallajosyula

Olive T. W. Li

Alex W. H. Chin

George Carnell

Nigel Temperton

Raghavan Varadarajan

Leo L. M. Poon

Crossref

Name not available

Antigenic drift of the HA head necessitates seasonal influenza vaccination, therefore in a bid to develop a universal vaccine, we adopted the approach of protein minimization of the HA protein to focus the antibody response towards the functionally conserved epitopes of the HA stem, using a H5 and H1 based vaccine in a mouse model. The H5 HA mini-stem protein refolds as a trimer, in a highly stable pre-fusion conformation representing the native conformation of the HA stem. The HA mini-stem is biochemically stable under diverse stress conditions, rendering the vaccine compatible for bulk production from E. Coli and pandemic stockpiling. Due to the ideal conformational nature of our HA mini-stem, antibodies elicited by the vaccine competed with other broadly neutralizing stem-specific antibodies (CR6261, F10 and FI6v3). Vaccine stem-specific antibodies were broadly reactive against HA proteins, binding with high affinity. Ultimately, vaccination of mice with the H1 and H5 HA mini-stem resulted in significant protection from morbidity and mortality against challenge with homologous and heterologous influenza viruses, including group 2 influenza, H3N2 challenge. Ours study is the first report of a Group 1 HA-stem vaccine showing Group 2 protection. Passive immune sera transfer demonstrated protection was mediated by vaccine stem-specific antibodies. Virus micro-neutralization was observed by vaccine immune sera for H1N1, H7N7 and H3N2 viruses, but not H5N1 viruses, despite clear protection in vaccinated and passive transfer experiments for H5N1 viruses. This raises important issues for the role of stem-specific antibodies, and whether neutralization is a requisite for board spectrum protection. We believe that this observation will provide critical research directions for developing novel universal influenza vaccines

http://hub.hku.hk/bitstream/10722/230500/1/content.pdf

Stalking influenza by vaccination with pre-fusion headless HA mini-stem.

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