Recombinant antigens prepared from the urease subunits of Helicobacter spp.: evidence of protection in a mouse model of gastric infection.
Urease is an important virulence factor for gastric Helicobacter spp. To elucidate the efficacy of individual urease subunits to act as mucosal immunogens, the genes encoding the respective urease subunits (UreA and UreB) of Helicobacter pylori and Helicobacter felis were cloned in an expression vector (pMAL) and expressed in Escherichia coli cells as translational fusion proteins. The recombinant UreA and UreB proteins were purified by affinity and anion-exchange chromatography techniques and had predicted molecular masses of approximately 68 and 103 kDa, respectively. Western blotting (immunoblotting) studies indicated that the urease components of the fusion proteins were strongly immunogenic and were specifically recognized by polyclonal rabbit anti-Helicobacter sp. sera. The fusion proteins (50 micrograms) were used, in combination with a mucosal adjuvant (cholera toxin), to orogastrically immunize mice against H. felis infection. Gastric tissues from H. felis-challenged mice were assessed by the biopsy urease test and by histology. In mice immunized with recombinant H. felis UreB, 60% of animals (n = 7) were histologically negative for H. felis bacteria after challenge at 17 weeks. This compared with 25% (n = 8) for mice immunized with the heterologous H. pylori UreB antigen. Neither the homologous nor the heterologous UreA subunit elicited protective responses against H. felis infection in mice. The study demonstrated that a recombinant subunit antigen could induce an immunoprotective response against gastric Helicobacter infection.
Ferrero
R L
RL
Unité des Entérobactéries (INSERM U389), Institut Pasteur, Paris, France.
Thiberge
J M
JM
Huerre
M
M
Labigne
A
A
D000818:Animals; D000942:Antigens, Bacterial / Q000276:immunology*; D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry; D005753:Gastric Mucosa / Q000276:immunology / Q000473:pathology; D016998:Helicobacter / Q000235:genetics / Q000276:immunology*; D016481:Helicobacter Infections / Q000276:immunology* / Q000473:pathology; D007111:Immunity, Cellular; D051379:Mice; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000276:immunology; D013997:Time Factors; D014510:Urease / Q000235:genetics / Q000276:immunology*
D000942:Antigens, Bacterial; D017931:DNA Primers; D011993:Recombinant Fusion Proteins; D014510:Urease