Expression of disulphide-bridge-dependent conformational epitopes and immunogenicity of the carboxy-terminal 19 kDa domain of Plasmodium yoelii merozoite surface protein-1 in live attenuated Salmonella vaccine strains.
The 19 kDa carboxy-terminal domain of Plasmodium yoelii merozoite surface protein-1 (MSP1(19)) was expressed in Salmonella vaccine strains as a carboxy-terminal fusion to fragment C of tetanus toxin (TetC). This study demonstrates that antibodies that recognize disulphide-dependent conformational epitopes in native MSP1 react with the TetC-MSP1(19) fusion protein expressed in Salmonella. The proper folding of MSP1(19) polypeptide is dependent on both the Salmonella host strain and the protein to which the MSP1(19) polypeptide is fused. Serum from mice immunized with Salmonella typhimurium C5aroD expressing TetC-MSP1(19) recognized native MSP1 as shown by immunofluorescence with P. yoelii-infected erythrocytes. Antibody levels to MSP1(19) were highest in out-bred mice immunized with S. typhimurium C5aroD carrying pTECH2-MSP1(19) and antibody was mostly directed against reduction-sensitive conformational epitopes. However, antibody levels were lower than in BALB/c mice immunized with a glutathione S-transferase (GST)-MSP1(19) fusion protein in Freund's adjuvant, and which were protected against P. yoelii challenge infection. In challenge experiments with P. yoelii the Salmonella-immunized mice were not protected, probably reflecting the magnitude of the antibody response. The results of this study have important implications in the design of live multivalent bacterial vaccines against eukaryotic pathogens.
Somner
Elizabeth A
EA
Department of Microbiology, The Medical School, University of Newcastle, Newcastle upon Tyne NE2 4HH, UK.
Ogun
Solabomi A
SA
Division of Parasitology, National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK.
Sinha
Katharine A
KA
Department of Microbiology, The Medical School, University of Newcastle, Newcastle upon Tyne NE2 4HH, UK.
Spencer Valero
Lilian M
LM
Division of Parasitology, National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK.
Lee
Jeong Jin
JJ
Department of Microbiology, The Medical School, University of Newcastle, Newcastle upon Tyne NE2 4HH, UK.
Harrison
Julia A
JA
Department of Microbiology, The Medical School, University of Newcastle, Newcastle upon Tyne NE2 4HH, UK.
Holder
Anthony A
AA
Division of Parasitology, National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK.
Hormaeche
Carlos E
CE
Department of Microbiology, The Medical School, University of Newcastle, Newcastle upon Tyne NE2 4HH, UK.
Khan
Anjam C M
ACM
Department of Microbiology, The Medical School, University of Newcastle, Newcastle upon Tyne NE2 4HH, UK.
D000818:Animals; D000913:Antibodies, Protozoan / Q000096:biosynthesis / Q000276:immunology*; D004220:Disulfides / Q000737:chemistry*; D000939:Epitopes / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D004912:Erythrocytes / Q000469:parasitology; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000235:genetics; D007114:Immunization; D008288:Malaria / Q000097:blood / Q000276:immunology / Q000517:prevention & control; D017780:Malaria Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D020066:Merozoite Surface Protein 1 / Q000097:blood / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics; D010967:Plasmodium yoelii / Q000235:genetics / Q000276:immunology*; D011487:Protein Conformation; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000276:immunology; D012475:Salmonella / Q000145:classification / Q000235:genetics* / Q000276:immunology; D013744:Tetanus Toxin / Q000096:biosynthesis / Q000235:genetics; D014613:Vaccines, Attenuated / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology
D000913:Antibodies, Protozoan; D004220:Disulfides; D000939:Epitopes; D017780:Malaria Vaccines; D020066:Merozoite Surface Protein 1; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D013744:Tetanus Toxin; D014613:Vaccines, Attenuated; D014614:Vaccines, Synthetic; C040635:tetanus toxin fragment C; D005982:Glutathione Transferase