title: Cell-penetrating peptides mediated protein cross-membrane delivery and its use in bacterial vector vaccine.
abstract: It is an attractive strategy to develop a recombinant bacterial vector vaccine by expressing exogenous protective antigen to induce the immune response, and the main concern is how to enhance the cellular internalization of antigen produced by bacterial vector. Cell-penetrating peptides (CPPs) are short cationic/amphipathic peptides which facilitate cellular uptake of various molecular cargoes and therefore have great potentials in vector vaccine design. In this work, eleven different CPPs were fused to the C-terminus of EGFP respectively, and the resultant EGFP-CPP fusion proteins were expressed and purified to assay their cross-membrane transport in macrophage J774 A.1 cells. Among the tested CPPs, TAT showed an excellent capability to deliver the cargo protein EGFP into cytoplasm. In order to establish an efficient antigen delivery system in Escherichia coli, the EGFP-TAT synthesis circuit was combined with an in vivo inducible lysis circuit PviuA-E in E. coli to form an integrated antigen delivery system, the resultant E. coli was proved to be able to lyse upon the induction of a mimic in vivo signal and thus release intracellular EGFP-TAT intensively, which were assumed to undergo a more efficient intracellular delivery by CPP to evoke protective immune responses. Based on the established antigen delivery system, the protective antigen gene flgD from an invasive intracellular fish pathogen Edwardsiella tarda EIB202, was applied to establish an E. coli recombinant vector vaccine. This E. coli vector vaccine presented superior immune protection (RPS = 63%) under the challenge with E. tarda EIB202, suggesting that the novel antigen delivery system had great potential in bacterial vector vaccine applications.
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, PR China.
authors.forename: Jimei
authors.initials: J
authors.lastname: Ma
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, PR China.
authors.forename: Jinmei
authors.initials: J
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, PR China.
authors.forename: Lingyu
authors.initials: L
authors.lastname: Guan
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, PR China.
authors.forename: Tianjian
authors.initials: T
authors.lastname: Hu
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, PR China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, PR China. Electronic address: jfxiao@ecust.edu.cn.
authors.forename: Jingfan
authors.initials: J
authors.lastname: Xiao
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, PR China.
authors.forename: Yuanxing
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D001428:Bacterial Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D001433:Bacteriolysis; D057846:Cell-Penetrating Peptides / Q000235:genetics* / Q000276:immunology; D020609:Edwardsiella tarda / Q000276:immunology*; D004756:Enterobacteriaceae Infections / Q000276:immunology / Q000662:veterinary*; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005393:Fish Diseases / Q000276:immunology*; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D011897:Random Allocation; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D015027:Zebrafish / Q000276:immunology*
chemical_list: D000942:Antigens, Bacterial; D001428:Bacterial Vaccines; D057846:Cell-Penetrating Peptides; D029968:Escherichia coli Proteins; D014614:Vaccines, Synthetic; C560213:lysis protein, E coli
keywords: Antigen delivery; Bacterial vector vaccine; Cell-penetrating peptides; Edwardsiella tarda