title: High-level expression of recombinant dengue virus type 2 envelope domain III protein and induction of neutralizing antibodies in BALB/C mice.
abstract: Dengue fever is a growing public health problem in many countries since so far no effective vaccines are available. In this study, the domain III of dengue virus type 2 envelope was expressed in Escherichia coli without fusion of any carrier protein. The recombinant protein was detected in the form of inclusion bodies, which were solubilized in 8M urea and could be purified subsequently by high-performance liquid chromatography (HPLC) on an ion exchange column. After refolding, the recombinant protein inhibited the DEN-2 plaque formation on C6/36 cells, demonstrated its function of receptor-interaction was retained. The recombinant protein was inoculated into BALB/c mice to test its immunogenicity and ability to induce neutralizing antibodies. The mice immunized with the purified protein developed high antibody titers. A neutralizing titer of 1:64 was also obtained by a cytopathogenic effect (CPE) inhibition assay in C6/36 cells. Mice challenged with lethal dose of DEN-2 in combination with sera from immunized mice were protected completely. The results suggested that these expression and purification strategies have the potential for development of an inexpensive vaccine.
authors.affiliation: Fujian Center for Disease Control and Prevention, Jintai Road 76, Fuzhou 35001, China.
authors.forename: Zhi-Shan
authors.initials: ZS
authors.lastname: Zhang
authors.forename: Yan-Sheng
authors.initials: YS
authors.lastname: Yan
authors.forename: Yu-Wei
authors.initials: YW
authors.lastname: Weng
authors.forename: Hai-Long
authors.initials: HL
authors.lastname: Huang
authors.forename: Shi-Qing
authors.initials: SQ
authors.lastname: Li
authors.forename: Shi
authors.initials: S
authors.lastname: He
authors.forename: Jian-Ming
authors.initials: JM
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D003588:Cytopathogenic Effect, Viral; D003715:Dengue / Q000517:prevention & control; D053059:Dengue Vaccines / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D003716:Dengue Virus / Q000276:immunology*; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D015870:Gene Expression; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D016019:Survival Analysis; D014614:Vaccines, Synthetic / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D010948:Viral Plaque Assay
chemical_list: D000914:Antibodies, Viral; D053059:Dengue Vaccines; C051821:E-glycoprotein, Dengue virus type 2; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins