Expression of a poliovirus type 1 neutralization epitope on a diphtheria toxin fusion protein.
The diphtheria toxin (DT) secreted by Corynebacterium diphtheriae is used after formolization as an efficient vaccine against diphtheria. In an attempt to evaluate its capacity to present heterologous peptide sequence in a recognized form, we created in-phase insertion in the gene encoding the non-toxic mutant protein CRM228 of DT. The sequence chosen for insertion was the synthetic DNA fragment encoding a poliovirus neutralization epitope. Tripartite fusion proteins comprising the mutant DT, the poliovirus peptide and beta-galactosidase were obtained in E. coli and purified by affinity chromatography. These fusion proteins reacted both with antibodies directed against the DT and a poliovirus specific monoclonal antibody. Moreover, these hybrid toxins induced protective antibodies against the lethal effect of DT and neutralizing antibodies against poliovirus. We conclude that the modification of highly immunogenic DT may provide a means for the presentation of foreign peptide sequences to the immune system.
Phalipon
A
A
Unité des Entérobactéries Institut Pasteur, Paris, France.
Crainic
R
R
Kaczorek
M
M
D000818:Animals; D015153:Blotting, Western; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004167:Diphtheria Toxin / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000276:immunology*; D008297:Male; D010957:Plasmids; D017955:Poliovirus / Q000276:immunology*; D011817:Rabbits; D014760:Viral Fusion Proteins / Q000032:analysis / Q000096:biosynthesis / Q000276:immunology*
D004167:Diphtheria Toxin; D000939:Epitopes; D014760:Viral Fusion Proteins