Cloning and Expression of aHelicobacter bilisImmunoreactive Protein

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Abstract
In an effort to identify immunoreactiveHelicobacter bilisantigens with potential for serodiagnosis, sera from mice experimentally infected withH. biliswere used to screen anH. bilisgenomic DNA expression library. Among 17 immunoreactive clones, several contained sequences that encoded a predicted 167-kDa protein (P167). Five overlapping P167 peptides (P167A to P167E) of approximately 40 kDa each were generated and tested. Immune sera reacted with fragments P167C and P167D at dilutions of 1:1,600 and 1:6,400, respectively, and reacted with anH. bilismembrane extract at a dilution of 1:800 in an enzyme-linked immunosorbent assay. Sera from mice experimentally infected withH. hepaticusdid not react with P167C and P167D. Sera from mice naturally infected withH. bilisbut not sera from mice naturally infected withH. hepaticusreacted with P167C and P167D. Hyperimmune sera against P167C peptide reacted with recombinant P167C and with a 120-kDa band inH. bilislysates but did not react with a protein of the same size on immunoblots prepared fromH. hepaticus,H. muridarum, or unrelatedBorrelia burgdorferiandCampylobacter jejuniwhole-cell lysates. Nevertheless, the P167A, P167B, P167C, and P167D primers, but not the P167E primers, amplified DNA fromH. hepaticus, and all five primer sets amplified DNA fromH. muridarum. These results suggest that P167 is an immunodominant,H. bilis-specific antigen that may have potential for use in serodiagnosis.

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