Construction, expression, and immunogenicity of multiple tandem copies of the Schistosoma mansoni peptide 115-131 of the P28 glutathione S-transferase expressed as C-terminal fusions to tetanus toxin fragment C in a live aro-attenuated vaccine strain of Salmonella.
Genetic fusions have been constructed between the highly immunogenic but atoxic fragment C of tetanus toxin and a guest peptide, aa115-131, from the protective 28-kDa glutathione S-transferase Ag of Schistosoma mansoni. Fusions have been assembled to express one, two, four, and eight tandem copies of the peptide. The recombinant vectors have been electroporated into the nonvirulent aroA strain of Salmonella typhimurium SL3261. The fusion proteins are soluble and stably expressed in Salmonella as evaluated by Western blotting with fragment C and glutathione S-transferase antisera. Mice have been immunized i.v. with a single dose of the live recombinant salmonellae. The strains are stable in mice and elicit Ab responses directed against fragment C, as determined by enzyme-linked immunosorbent assays. Ab responses were also detected against the guest peptide. The Ab responses improved dramatically toward the aa115-131 peptide with increasing copy number, with the octameric "repitope" fusion displaying the greatest potency. This approach may represent a general strategy for eliciting immune responses against peptides in live bacterial vaccines.
Khan
C M
CM
Department of Pathology, University of Cambridge, UK.
Villarreal-Ramos
B
B
Pierce
R J
RJ
Demarco de Hormaeche
R
R
McNeill
H
H
Ali
T
T
Chatfield
S
S
Capron
A
A
Dougan
G
G
Hormaeche
C E
CE
D000595:Amino Acid Sequence; D000818:Animals; D001428:Bacterial Vaccines; D001483:Base Sequence; D015153:Blotting, Western; D000939:Epitopes / Q000276:immunology; D005260:Female; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000276:immunology*; D015801:Helminth Proteins / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000276:immunology*; D012486:Salmonella typhimurium / Q000276:immunology; D012550:Schistosoma mansoni / Q000276:immunology*; D013744:Tetanus Toxin / Q000276:immunology; D014613:Vaccines, Attenuated
D001428:Bacterial Vaccines; D000939:Epitopes; D015801:Helminth Proteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D013744:Tetanus Toxin; D014613:Vaccines, Attenuated; C040635:tetanus toxin fragment C; D005982:Glutathione Transferase