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Pore-forming alpha-hemolysin efficiently improves the immunogenicity and protective efficacy of protein antigens
Highly immunogenic exotoxins are used as carrier proteins because they efficiently improve the immunogenicity of polysaccharides. However, their efficiency with protein antigens remains unclear. In the current study, the candidate antigen PA0833 from Pseudomonas aeruginosa was fused to the α-hemolys...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8294524/ https://www.ncbi.nlm.nih.gov/pubmed/34288976 http://dx.doi.org/10.1371/journal.ppat.1009752 |
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author | Zou, Jin-Tao Jing, Hai-Ming Yuan, Yue Lei, Lang-Huan Chen, Zhi-Fu Gou, Qiang Xiong, Qing-Shan Zhang, Xiao-Li Zhao, Zhuo Zhang, Xiao-Kai Zeng, Hao Zou, Quan-Ming Zhang, Jin-Yong |
author_facet | Zou, Jin-Tao Jing, Hai-Ming Yuan, Yue Lei, Lang-Huan Chen, Zhi-Fu Gou, Qiang Xiong, Qing-Shan Zhang, Xiao-Li Zhao, Zhuo Zhang, Xiao-Kai Zeng, Hao Zou, Quan-Ming Zhang, Jin-Yong |
author_sort | Zou, Jin-Tao |
collection | PubMed |
description | Highly immunogenic exotoxins are used as carrier proteins because they efficiently improve the immunogenicity of polysaccharides. However, their efficiency with protein antigens remains unclear. In the current study, the candidate antigen PA0833 from Pseudomonas aeruginosa was fused to the α-hemolysin mutant Hla(H35A) from Staphylococcus aureus to form a Hla(H35A)-PA0833 fusion protein (HPF). Immunization with HPF resulted in increased PA0833-specific antibody titers, higher protective efficacy, and decreased bacterial burden and pro-inflammatory cytokine secretion compared with PA0833 immunization alone. Using fluorescently labeled antigens to track antigen uptake and delivery, we found that Hla(H35A) fusion significantly improved antigen uptake in injected muscles and antigen delivery to draining lymph nodes. Both in vivo and in vitro studies demonstrated that the increased antigen uptake after immunization with HPF was mainly due to monocyte- and macrophage-dependent macropinocytosis, which was probably the result of HPF binding to ADAM10, the Hla host receptor. Furthermore, a transcriptome analysis showed that several immune signaling pathways were activated by HPF, shedding light on the mechanism whereby Hla(H35A) fusion improves immunogenicity. Finally, the improvement in immunogenicity by Hla(H35A) fusion was also confirmed with two other antigens, GlnH from Klebsiella pneumoniae and the model antigen OVA, indicating that Hla(H35A) could serve as a universal carrier protein to improve the immunogenicity of protein antigens. |
format | Online Article Text |
id | pubmed-8294524 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-82945242021-07-31 Pore-forming alpha-hemolysin efficiently improves the immunogenicity and protective efficacy of protein antigens Zou, Jin-Tao Jing, Hai-Ming Yuan, Yue Lei, Lang-Huan Chen, Zhi-Fu Gou, Qiang Xiong, Qing-Shan Zhang, Xiao-Li Zhao, Zhuo Zhang, Xiao-Kai Zeng, Hao Zou, Quan-Ming Zhang, Jin-Yong PLoS Pathog Research Article Highly immunogenic exotoxins are used as carrier proteins because they efficiently improve the immunogenicity of polysaccharides. However, their efficiency with protein antigens remains unclear. In the current study, the candidate antigen PA0833 from Pseudomonas aeruginosa was fused to the α-hemolysin mutant Hla(H35A) from Staphylococcus aureus to form a Hla(H35A)-PA0833 fusion protein (HPF). Immunization with HPF resulted in increased PA0833-specific antibody titers, higher protective efficacy, and decreased bacterial burden and pro-inflammatory cytokine secretion compared with PA0833 immunization alone. Using fluorescently labeled antigens to track antigen uptake and delivery, we found that Hla(H35A) fusion significantly improved antigen uptake in injected muscles and antigen delivery to draining lymph nodes. Both in vivo and in vitro studies demonstrated that the increased antigen uptake after immunization with HPF was mainly due to monocyte- and macrophage-dependent macropinocytosis, which was probably the result of HPF binding to ADAM10, the Hla host receptor. Furthermore, a transcriptome analysis showed that several immune signaling pathways were activated by HPF, shedding light on the mechanism whereby Hla(H35A) fusion improves immunogenicity. Finally, the improvement in immunogenicity by Hla(H35A) fusion was also confirmed with two other antigens, GlnH from Klebsiella pneumoniae and the model antigen OVA, indicating that Hla(H35A) could serve as a universal carrier protein to improve the immunogenicity of protein antigens. Public Library of Science 2021-07-21 /pmc/articles/PMC8294524/ /pubmed/34288976 http://dx.doi.org/10.1371/journal.ppat.1009752 Text en © 2021 Zou et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Zou, Jin-Tao Jing, Hai-Ming Yuan, Yue Lei, Lang-Huan Chen, Zhi-Fu Gou, Qiang Xiong, Qing-Shan Zhang, Xiao-Li Zhao, Zhuo Zhang, Xiao-Kai Zeng, Hao Zou, Quan-Ming Zhang, Jin-Yong Pore-forming alpha-hemolysin efficiently improves the immunogenicity and protective efficacy of protein antigens |
title | Pore-forming alpha-hemolysin efficiently improves the immunogenicity and protective efficacy of protein antigens |
title_full | Pore-forming alpha-hemolysin efficiently improves the immunogenicity and protective efficacy of protein antigens |
title_fullStr | Pore-forming alpha-hemolysin efficiently improves the immunogenicity and protective efficacy of protein antigens |
title_full_unstemmed | Pore-forming alpha-hemolysin efficiently improves the immunogenicity and protective efficacy of protein antigens |
title_short | Pore-forming alpha-hemolysin efficiently improves the immunogenicity and protective efficacy of protein antigens |
title_sort | pore-forming alpha-hemolysin efficiently improves the immunogenicity and protective efficacy of protein antigens |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8294524/ https://www.ncbi.nlm.nih.gov/pubmed/34288976 http://dx.doi.org/10.1371/journal.ppat.1009752 |
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