A DNA vaccine expressing consensus hemagglutinin-esterase fusion protein protected guinea pigs from infection by two lineages of influenza D virus

Yanmin Wan; Guobin Kang; Chithra Sreenivasan; Lance Daharsh; Junfeng Zhang; Wenjin Fan; Dan Wang; Hideaki Moriyama; Feng Li; Qingsheng Li

doi:10.1128/JVI.00110-18

A DNA vaccine expressing consensus hemagglutinin-esterase fusion protein protected guinea pigs from infection by two lineages of influenza D virus

Yanmin Wan, Guobin Kang, Chithra Sreenivasan, Lance Daharsh, Junfeng Zhang, Wenjin Fan, Dan Wang, Hideaki Moriyama, Feng Li, Qingsheng Li

Research output: Contribution to journal › Article › peer-review

11 Scopus citations

Abstract

Two lineages of influenza D virus (IDV) have been found to infect cattle and promote bovine respiratory disease complex, one of the most commonly diagnosed causes of morbidity and mortality within the cattle industry. Furthermore, IDV can infect other economically important domestic livestock, including pigs, and has the potential to infect humans, which necessitates the need for an efficacious vaccine. In this study, we designed a DNA vaccine expressing consensus hemagglutinin-esterase fusion (HEF) protein (FluD-Vax) and tested its protective efficacy against two lineages of IDV (D/OK and D/660) in guinea pigs. Animals that received FluD-Vax (n = 12) developed appreciable titers of neutralizing antibodies against IDV lineage representatives, D/OK and D/660. Importantly, vaccinated animals were protected against intranasal challenge with IDV [3 × 10⁵ 50% tissue culture infective dose(s) (TCID₅₀)] D/OK (n = 6) or D/600 (n = 6), based on the absence of viral RNA in necropsied tissues (5 and 7 days postchallenge) using quantitative reverse transcription-PCR and in situ hybridization. In contrast, animals that received a sham DNA vaccine (n = 12) had no detectable neutralizing antibodies against IDV, and viral RNA was readily detectable in respiratory tract tissues after intranasal challenge (3 × 10⁵ TCID₅₀) with IDV D/OK (n = 6) or D/660 (n = 6). Using a TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling) assay, we found that IDV D/OK and D/600 infections induced apoptosis in epithelial cells lining alveoli and bronchioles, as well as nonepithelial cells in lung tissues. Our results demonstrate for the first time that the consensus IDV HEF DNA vaccine can elicit complete protection against infection from two lineages of IDV in the guinea pig model.

Original language	English
Article number	e00110-18
Journal	Journal of Virology
Volume	92
Issue number	11
DOIs	https://doi.org/10.1128/JVI.00110-18
State	Published - Jun 1 2018

Bibliographical note

Funding Information:
Q.L. and Y.W. conceived the idea and designed the overall experiments with F.L. Y.W. designed and prepared the vaccine. Y.W., G.K., L.D., J.Z., and W.F. conducted the animal experiments. C.S. and D.W. prepared the virus stocks and measured the HI antibody titers. G.K. and J.F. conducted the ISH. Y.W. conducted the qRT-PCR. H.M. did the structural modeling of HEF protein receptor sites. Y.W. and Q.L. wrote the manuscript, and all authors commented and made revisions. We thank Shan Lu for providing pJW4303 expression vector and staff members at the Institutional Animal Care Program of UNL for assistance in animal care. This project was funded in part by start-up funds from UNL, grants SDSU AES 3AH-477 and NIH R21AI107379, National Science Foundation/EPSCoR (http://www.nsf .gov/od/iia/programs/epscor/index.jsp) award IIA-1335423, and the state of South Dakota Governor's Office of Economic Development as a South Dakota Research Innovation Center.

Funding Information:
This project was funded in part by start-up funds from UNL, grants SDSU AES 3AH-477 and NIH R21AI107379, National Science Foundation/EPSCoR (http://www.nsf .gov/od/iia/programs/epscor/index.jsp) award IIA-1335423, and the state of South Dakota Governor’s Office of Economic Development as a South Dakota Research Innovation Center.

Publisher Copyright:
© 2018 American Society for Microbiology.

Keywords

Consensus sequence
DNA vaccine
Guinea pigs
Hemagglutinin-esterase-fusion protein
Influenza D virus

ASJC Scopus subject areas

Microbiology
Immunology
Insect Science
Virology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1128/JVI.00110-18

Cite this

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title = "A DNA vaccine expressing consensus hemagglutinin-esterase fusion protein protected guinea pigs from infection by two lineages of influenza D virus",

abstract = "Two lineages of influenza D virus (IDV) have been found to infect cattle and promote bovine respiratory disease complex, one of the most commonly diagnosed causes of morbidity and mortality within the cattle industry. Furthermore, IDV can infect other economically important domestic livestock, including pigs, and has the potential to infect humans, which necessitates the need for an efficacious vaccine. In this study, we designed a DNA vaccine expressing consensus hemagglutinin-esterase fusion (HEF) protein (FluD-Vax) and tested its protective efficacy against two lineages of IDV (D/OK and D/660) in guinea pigs. Animals that received FluD-Vax (n = 12) developed appreciable titers of neutralizing antibodies against IDV lineage representatives, D/OK and D/660. Importantly, vaccinated animals were protected against intranasal challenge with IDV [3 × 105 50% tissue culture infective dose(s) (TCID50)] D/OK (n = 6) or D/600 (n = 6), based on the absence of viral RNA in necropsied tissues (5 and 7 days postchallenge) using quantitative reverse transcription-PCR and in situ hybridization. In contrast, animals that received a sham DNA vaccine (n = 12) had no detectable neutralizing antibodies against IDV, and viral RNA was readily detectable in respiratory tract tissues after intranasal challenge (3 × 105 TCID50) with IDV D/OK (n = 6) or D/660 (n = 6). Using a TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling) assay, we found that IDV D/OK and D/600 infections induced apoptosis in epithelial cells lining alveoli and bronchioles, as well as nonepithelial cells in lung tissues. Our results demonstrate for the first time that the consensus IDV HEF DNA vaccine can elicit complete protection against infection from two lineages of IDV in the guinea pig model.",

keywords = "Consensus sequence, DNA vaccine, Guinea pigs, Hemagglutinin-esterase-fusion protein, Influenza D virus",

author = "Yanmin Wan and Guobin Kang and Chithra Sreenivasan and Lance Daharsh and Junfeng Zhang and Wenjin Fan and Dan Wang and Hideaki Moriyama and Feng Li and Qingsheng Li",

note = "Funding Information: Q.L. and Y.W. conceived the idea and designed the overall experiments with F.L. Y.W. designed and prepared the vaccine. Y.W., G.K., L.D., J.Z., and W.F. conducted the animal experiments. C.S. and D.W. prepared the virus stocks and measured the HI antibody titers. G.K. and J.F. conducted the ISH. Y.W. conducted the qRT-PCR. H.M. did the structural modeling of HEF protein receptor sites. Y.W. and Q.L. wrote the manuscript, and all authors commented and made revisions. We thank Shan Lu for providing pJW4303 expression vector and staff members at the Institutional Animal Care Program of UNL for assistance in animal care. This project was funded in part by start-up funds from UNL, grants SDSU AES 3AH-477 and NIH R21AI107379, National Science Foundation/EPSCoR (http://www.nsf .gov/od/iia/programs/epscor/index.jsp) award IIA-1335423, and the state of South Dakota Governor's Office of Economic Development as a South Dakota Research Innovation Center. Funding Information: This project was funded in part by start-up funds from UNL, grants SDSU AES 3AH-477 and NIH R21AI107379, National Science Foundation/EPSCoR (http://www.nsf .gov/od/iia/programs/epscor/index.jsp) award IIA-1335423, and the state of South Dakota Governor{\textquoteright}s Office of Economic Development as a South Dakota Research Innovation Center. Publisher Copyright: {\textcopyright} 2018 American Society for Microbiology.",

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T1 - A DNA vaccine expressing consensus hemagglutinin-esterase fusion protein protected guinea pigs from infection by two lineages of influenza D virus

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AU - Daharsh, Lance

AU - Zhang, Junfeng

AU - Fan, Wenjin

AU - Wang, Dan

AU - Moriyama, Hideaki

AU - Li, Feng

AU - Li, Qingsheng

N1 - Funding Information: Q.L. and Y.W. conceived the idea and designed the overall experiments with F.L. Y.W. designed and prepared the vaccine. Y.W., G.K., L.D., J.Z., and W.F. conducted the animal experiments. C.S. and D.W. prepared the virus stocks and measured the HI antibody titers. G.K. and J.F. conducted the ISH. Y.W. conducted the qRT-PCR. H.M. did the structural modeling of HEF protein receptor sites. Y.W. and Q.L. wrote the manuscript, and all authors commented and made revisions. We thank Shan Lu for providing pJW4303 expression vector and staff members at the Institutional Animal Care Program of UNL for assistance in animal care. This project was funded in part by start-up funds from UNL, grants SDSU AES 3AH-477 and NIH R21AI107379, National Science Foundation/EPSCoR (http://www.nsf .gov/od/iia/programs/epscor/index.jsp) award IIA-1335423, and the state of South Dakota Governor's Office of Economic Development as a South Dakota Research Innovation Center. Funding Information: This project was funded in part by start-up funds from UNL, grants SDSU AES 3AH-477 and NIH R21AI107379, National Science Foundation/EPSCoR (http://www.nsf .gov/od/iia/programs/epscor/index.jsp) award IIA-1335423, and the state of South Dakota Governor’s Office of Economic Development as a South Dakota Research Innovation Center. Publisher Copyright: © 2018 American Society for Microbiology.

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N2 - Two lineages of influenza D virus (IDV) have been found to infect cattle and promote bovine respiratory disease complex, one of the most commonly diagnosed causes of morbidity and mortality within the cattle industry. Furthermore, IDV can infect other economically important domestic livestock, including pigs, and has the potential to infect humans, which necessitates the need for an efficacious vaccine. In this study, we designed a DNA vaccine expressing consensus hemagglutinin-esterase fusion (HEF) protein (FluD-Vax) and tested its protective efficacy against two lineages of IDV (D/OK and D/660) in guinea pigs. Animals that received FluD-Vax (n = 12) developed appreciable titers of neutralizing antibodies against IDV lineage representatives, D/OK and D/660. Importantly, vaccinated animals were protected against intranasal challenge with IDV [3 × 105 50% tissue culture infective dose(s) (TCID50)] D/OK (n = 6) or D/600 (n = 6), based on the absence of viral RNA in necropsied tissues (5 and 7 days postchallenge) using quantitative reverse transcription-PCR and in situ hybridization. In contrast, animals that received a sham DNA vaccine (n = 12) had no detectable neutralizing antibodies against IDV, and viral RNA was readily detectable in respiratory tract tissues after intranasal challenge (3 × 105 TCID50) with IDV D/OK (n = 6) or D/660 (n = 6). Using a TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling) assay, we found that IDV D/OK and D/600 infections induced apoptosis in epithelial cells lining alveoli and bronchioles, as well as nonepithelial cells in lung tissues. Our results demonstrate for the first time that the consensus IDV HEF DNA vaccine can elicit complete protection against infection from two lineages of IDV in the guinea pig model.

AB - Two lineages of influenza D virus (IDV) have been found to infect cattle and promote bovine respiratory disease complex, one of the most commonly diagnosed causes of morbidity and mortality within the cattle industry. Furthermore, IDV can infect other economically important domestic livestock, including pigs, and has the potential to infect humans, which necessitates the need for an efficacious vaccine. In this study, we designed a DNA vaccine expressing consensus hemagglutinin-esterase fusion (HEF) protein (FluD-Vax) and tested its protective efficacy against two lineages of IDV (D/OK and D/660) in guinea pigs. Animals that received FluD-Vax (n = 12) developed appreciable titers of neutralizing antibodies against IDV lineage representatives, D/OK and D/660. Importantly, vaccinated animals were protected against intranasal challenge with IDV [3 × 105 50% tissue culture infective dose(s) (TCID50)] D/OK (n = 6) or D/600 (n = 6), based on the absence of viral RNA in necropsied tissues (5 and 7 days postchallenge) using quantitative reverse transcription-PCR and in situ hybridization. In contrast, animals that received a sham DNA vaccine (n = 12) had no detectable neutralizing antibodies against IDV, and viral RNA was readily detectable in respiratory tract tissues after intranasal challenge (3 × 105 TCID50) with IDV D/OK (n = 6) or D/660 (n = 6). Using a TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling) assay, we found that IDV D/OK and D/600 infections induced apoptosis in epithelial cells lining alveoli and bronchioles, as well as nonepithelial cells in lung tissues. Our results demonstrate for the first time that the consensus IDV HEF DNA vaccine can elicit complete protection against infection from two lineages of IDV in the guinea pig model.

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KW - Hemagglutinin-esterase-fusion protein

KW - Influenza D virus

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A DNA vaccine expressing consensus hemagglutinin-esterase fusion protein protected guinea pigs from infection by two lineages of influenza D virus

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

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