[Construction, expression and immunogenicity analysis of a fusion protein containing M2e of influenza A virus fused to a modified Pseudomonas aeruginosa exotoxin A].
M2 protein of type A influenza virus is a good candidate for universal influenza vaccine, exotoxin A of Pseudomonas aeruginosa may facilitate the immunogenicity of M2 protein. We constructed and expressed a prokaryotic expression plasmid containing a chimeric gene of M2 extracellular coding region and a partial PEA gene, and observed the immunoprotection in BALB/c mice vaccinated with the fusion protein. The fusion protein (ntPE-M2e) was generated by inserting the coding sequence of the M2e in place of Ib loop in PEA. This fusion protein was used to immunize BALB/c mice by subcutaneously injection with incomplete Freund's adjuvant and boost at weeks 3 and 7. The immunized mice were challenged with influenza virus strain A/PR/34/8. The fusion protein (ntPE-M2e) immunization protected mice against lethal viral challenge. ELISA and ELISPOT results demonstrated that the fusion protein could induce a strong systemic immune response against synthetic M2e peptide, and virus replication in the lungs of mice was inhibited in comparison with the control. This study provides foundation for developing broad-spectrum vaccines against type A influenza viruses.
Xu
Yi
Y
National Key Laboratory of Molecular Virology and Genetic Engineering, Institute of Viral Disease Control and Prevention, China CDC, Beijing 100052, China.
Yao
Li-hong
LH
Chen
Ai-jun
AJ
Guo
Jian-qiang
JQ
Liu
Xiao-yu
XY
Bo
Hong
H
Liu
Li-qi
LQ
Shu
Yue-long
YL
Zhang
Zhi-qing
ZQ
D036002:ADP Ribose Transferases / Q000235:genetics*; D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005098:Exotoxins / Q000235:genetics*; D005260:Female; D015870:Gene Expression; D007114:Immunization; D009980:Influenza A virus / Q000276:immunology / Q000502:physiology; D008168:Lung / Q000276:immunology / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D014763:Viral Matrix Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D037521:Virulence Factors / Q000235:genetics*; D000097668:Pseudomonas aeruginosa Exotoxin A; D000086082:Viroporin Proteins
D036002:ADP Ribose Transferases; D001427:Bacterial Toxins; D005098:Exotoxins; D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins