title: Recombinant tandem multi-linear neutralizing epitopes of human enterovirus 71 elicited protective immunity in mice.
abstract: BACKGROUND
  Human Enterovirus 71 (EV71) has emerged as the leading cause of viral encephalitis in children, especially in the Asia-Pacific regions. EV71 vaccine development is of high priority at present, and neutralization antibodies have been documented to play critical roles during in vitro and in vivo protection against EV71 infection.
  
  
  RESULTS
  In this study, a novel strategy to produce EV71 vaccine candidate based on recombinant multiple tandem linear neutralizing epitopes (mTLNE) was proposed. The three well identified EV71 linear neutralizing epitopes in capsid proteins, VP1-SP55, VP1-SP70 and VP2-SP28, were sequentially linked by a Gly-Ser linker ((G4S)3), and expressed in E.coli in fusion with the Trx and His tag at either terminal. The recombinant protein mTLNE was soluble and could be purified by standard affinity chromatography. Following three dosage of immunization in adult mice, EV71-specific IgG and neutralization antibodies were readily induced by recombinant mTLNE. IgG subtyping demonstrated that lgG1 antibodies dominated the mTLNE-induced humoral immune response. Especially, cytokine profiling in spleen cells from the mTLNE-immunized mice revealed high production of IL-4 and IL-6. Finally, in vivo challenge experiments showed that passive transfer with anti-mTLNE sera conferred full protection against lethal EV71 challenge in neonatal mice.
  
  
  CONCLUSIONS
  Our results demonstrated that this rational designed recombinant mTLNE might have the potential to be further developed as an EV71 vaccine in the future.
authors.forename: Yue-Xiang
authors.initials: YX
authors.lastname: Li
authors.forename: Hui
authors.initials: H
authors.lastname: Zhao
authors.forename: Rui-Yuan
authors.initials: RY
authors.lastname: Cao
authors.forename: Yong-Qiang
authors.initials: YQ
authors.lastname: Deng
authors.forename: Jian-Feng
authors.initials: JF
authors.lastname: Han
authors.forename: Shun-Ya
authors.initials: SY
authors.lastname: Zhu
authors.forename: Jie
authors.initials: J
authors.lastname: Ma
authors.forename: Long
authors.initials: L
authors.lastname: Liu
authors.forename: E-De
authors.initials: ED
authors.lastname: Qin
authors.affiliation: Graduate School, Anhui Medical University, Hefei 230032, Anhui, China. qincf@bmi.ac.cn.
authors.forename: Cheng-Feng
authors.initials: CF
authors.lastname: Qin
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology*; D002846:Chromatography, Affinity; D016207:Cytokines / Q000032:analysis; D004195:Disease Models, Animal; D029821:Enterovirus A, Human / Q000276:immunology*; D004769:Enterovirus Infections / Q000276:immunology / Q000517:prevention & control*; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D015870:Gene Expression; D007116:Immunization, Passive; D007074:Immunoglobulin G / Q000097:blood; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology*; D016019:Survival Analysis; D014611:Vaccination / Q000379:methods; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014765:Viral Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D016207:Cytokines; D018985:Epitopes, B-Lymphocyte; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D014765:Viral Vaccines