title: Establishment of a novel method without sequence modification for developing NoV P particle-based chimeric vaccines.
abstract: The Norovirus (NoV) P particle (PP) is a subviral particle formed by 24 copies of the protruding (P) domain of the capsid protein. Each P domain has three surface loops that can be used for foreign antigen presentation. Hence, PPs have been demonstrated to be an excellent platform for vaccine development against many pathogens. However, current processes for preparing those chimeric PP vaccines vary and would change the original sequence of the PP. A detailed strategy also has not been reported for inserting a foreign antigen into all three loops. In order to develop a novel method for preparing distinct types of PP-based protein vaccines, we created two restriction enzyme sites (EagI and KpnI) in the P domain by site-directed mutagenesis without changing its original sequence. A synthesized gene with three copies of the Alzheimer's disease (AD) immunogen Aβ1-6 was then incorporated in loop2 of the P domain. Additionally, a synthesized gene with one copy of Aβ1-6 was inserted into each loop of the P domain. Furthermore, two recombinant proteins PP-3 copy-Aβ1-6-loop2 and PP-1 copy-Aβ1-6-loop123 were successfully purified without affecting PP formation. Particle size analysis and TEM observations demonstrated that the two chimeric P particles were still able to form 24-mer nanoparticles. Moreover, the two chimeric PP-based AD vaccines could both efficiently elicit strong immune responses in the mouse model. In conclusion, we have successfully established a novel method for preparing vaccines based on the NoV PP which would not affect PP sequence and function.
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun 130012, China.
authors.forename: Yingnan
authors.initials: Y
authors.lastname: Li
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun 130012, China; Key Laboratory for Molecular Enzymology and Engineering, The Ministry of Education, School of Life Sciences, Jilin University, Changchun 130012, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Fu
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun 130012, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Hu
authors.affiliation: School of Chemistry and Life Science, Changchun University of Technology, Changchun 130012, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Jin
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun 130012, China.
authors.forename: Yayuan
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: School of Chemistry and Life Science, Changchun University of Technology, Changchun 130012, China.
authors.forename: Yuhe
authors.initials: Y
authors.lastname: Yin
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun 130012, China; Key Laboratory for Molecular Enzymology and Engineering, The Ministry of Education, School of Life Sciences, Jilin University, Changchun 130012, China. Electronic address: topwuhui@jlu.edu.cn.
authors.forename: Hui
authors.initials: H
authors.lastname: Wu
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun 130012, China; Key Laboratory for Molecular Enzymology and Engineering, The Ministry of Education, School of Life Sciences, Jilin University, Changchun 130012, China. Electronic address: xianghui@jlu.edu.cn.
authors.forename: Xianghui
authors.initials: X
authors.lastname: Yu
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun 130012, China; Key Laboratory for Molecular Enzymology and Engineering, The Ministry of Education, School of Life Sciences, Jilin University, Changchun 130012, China. Electronic address: weikong@jlu.edu.cn.
authors.forename: Wei
authors.initials: W
authors.lastname: Kong
mesh_terms: D000544:Alzheimer Disease / Q000276:immunology* / Q000517:prevention & control; D000818:Animals; D017951:Antigen Presentation / Q000276:immunology; D036022:Capsid Proteins / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology; D006801:Humans; D051379:Mice; D016297:Mutagenesis, Site-Directed; D053758:Nanoparticles / Q000008:administration & dosage; D029322:Norovirus / Q000235:genetics / Q000276:immunology*; D058425:Vaccines, Virus-Like Particle / Q000008:administration & dosage / Q000276:immunology*; D014765:Viral Vaccines / Q000235:genetics / Q000276:immunology* / Q000627:therapeutic use
chemical_list: D036022:Capsid Proteins; D058425:Vaccines, Virus-Like Particle; D014765:Viral Vaccines
keywords: AD; Aβ1-6; Chimeric vaccine; Noroviruses; P particle