Isolation of recombinant protective Helicobacter pylori antigens.
A total of seven clones producing both new and previously described Helicobacter pylori proteins were isolated from a library of H. pylori genomic DNA. The screening approach by which these proteins were detected relied on the use of antisera raised in mice vaccinated with Helicobacter felis sonicate plus cholera toxin, a regimen which protects mice from H. pylori challenge. This strategy was designed to maximize the possibility of obtaining antigens which might be capable of conferring protection from H. pylori infection. Two of the clones were shown to encode the urease enzyme and the heat shock protein HspB, which have already been identified as protective antigens. The other five clones were sequenced, protein coding regions were deduced, and these sequences were amplified by PCR for incorporation into Escherichia coli expression vectors. The proteins produced from these expression systems were purified to allow testing for protective efficacy in an H. pylori mouse model. All five proteins were able to facilitate the clearance of a challenge with H. pylori, as judged by an assay of gastric urease activity and light microscopy on stomach sections. These results clearly indicate that the screening strategy has successfully identified candidate vaccine antigens.
Hocking
D
D
Research and Development Division, CSL Limited, Parkville, Victoria, Australia 3052.
Webb
E
E
Radcliff
F
F
Rothel
L
L
Taylor
S
S
Pinczower
G
G
Kapouleas
C
C
Braley
H
H
Lee
A
A
Doidge
C
C
D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D001483:Base Sequence; D004269:DNA, Bacterial; D015870:Gene Expression; D016680:Genome, Bacterial; D015698:Genomic Library; D016481:Helicobacter Infections / Q000517:prevention & control; D016480:Helicobacter pylori / Q000235:genetics* / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA; D014510:Urease / Q000235:genetics
D000942:Antigens, Bacterial; D004269:DNA, Bacterial; D014510:Urease