Recombinant protein comprising multi-neutralizing epitopes induced high titer of antibodies against influenza A virus.
In previous studies, we suggested that epitope-vaccine might be a new strategy against virus infection. Based on this hypothesis, we designed and expressed a recombinant immunogen (multi-epitope-peptide) comprising repeats of three neutralizing-epitopes (neutralizing epitopes: aa92-105, 127-133 and 183-195) of hemagglutininin (HA) of influenza virus (H3N2) in E. coli. After vaccination, the recombinant multi-epitope protein could induce a high level of antibodies with predefined multi-epitope-specificity in mice and rabbits. The epitope-specific antibodies in sera were tested using three different epitope-peptides (synthetic peptides) in ELISA assay, and the serum dilutions from 1 : 6400 to 1 : 25600 were confirmed. In western blot analysis, both the antiserum and the antibodies purified by synthetic epitope-peptide coupled sepharose columns could recognize natural HA from influenza virus particles (strain A/Wuhan/359/95 H3N2). In hemagglutination inhibition (HI) tests, these three antisera at the dilutions from 1 : 20 to 1 : 80 showed inhibitory activity. Interestingly, antisera and purified antibodies induced by the epitope-vaccine could partially inhibit plaque-formation of influenza virus (strain A/Wuhan/359/95) on MDCK cell monolayers. These results suggest that the recombinant multi-epitope vaccine can simultaneously induce multi-antiviral activities against influenza virus, which may provide a new way to develop effective vaccines against influenza virus.
Laboratory of Immunology, Research Centre for Medical Science, Department of Biology, Tsinghua University, Beijing 100084, P R China.
Hua
H
Li
Jian
J
Ding
Ying-Hua
YH
Chen
D000818:Animals; D000906:Antibodies / Q000097:blood / Q000276:immunology*; D001483:Base Sequence; D002460:Cell Line; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D006385:Hemagglutination Inhibition Tests; D006388:Hemagglutinins / Q000235:genetics / Q000276:immunology*; D009980:Influenza A virus / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D009976:Orthomyxoviridae Infections / Q000276:immunology* / Q000821:virology*; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D010948:Viral Plaque Assay; D014765:Viral Vaccines / Q000097:blood / Q000235:genetics / Q000276:immunology*
D000906:Antibodies; D000939:Epitopes; D006388:Hemagglutinins; D011994:Recombinant Proteins; D014765:Viral Vaccines