title: Molecular and immunological characterization of the highly conserved antigen 84 from Mycobacterium tuberculosis and Mycobacterium leprae.
abstract: Crossed immunoelectrophoresis (CIE) has been used to develop a reference system for classifying mycobacterial antigens. The subsequent use of specific antibodies allowed further determination of antigens by molecular weight. The monoclonal antibody F126-2, originally raised against a 34-kDa antigen of Mycobacterium kansasii, reacted with antigen 84 (Ag84) in the CIE reference system for Mycobacterium bovis BCG and Mycobacterium tuberculosis. To characterize Ag84, we screened a lambda gt11 gene library from M. tuberculosis with antibody F126-2 and identified the encoding gene. The corresponding Mycobacterium leprae Ag84 gene was subsequently selected from a cosmid library, using the M. tuberculosis gene as a probe. Both genes were expressed as 34-kDa proteins in Escherichia coli, and the recombinant proteins indeed corresponded to Ag84 in the CIE reference system. The derived amino acid sequences of the M. tuberculosis and M. leprae proteins showed 85% identity, which indicates that Ag84 constitutes a group of highly conserved mycobacterial antigens. Antibodies of almost 60% of lepromatous leprosy patients responded to Ag84, indicating that the protein is highly immunogenic following infection in multibacillary leprosy.
authors.affiliation: Armauer Hansen Research Institute, Addis Ababa, Ethiopia.
authors.forename: P W
authors.initials: PW
authors.lastname: Hermans
authors.forename: F
authors.initials: F
authors.lastname: Abebe
authors.forename: V I
authors.initials: VI
authors.lastname: Kuteyi
authors.forename: A H
authors.initials: AH
authors.lastname: Kolk
authors.forename: J E
authors.initials: JE
authors.lastname: Thole
authors.forename: M
authors.initials: M
authors.lastname: Harboe
mesh_terms: D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000096:biosynthesis / Q000276:immunology; D000942:Antigens, Bacterial / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D017124:Conserved Sequence; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D005798:Genes, Bacterial / Q000235:genetics*; D006801:Humans; D007123:Immunoelectrophoresis, Two-Dimensional; D007918:Leprosy / Q000097:blood; D008969:Molecular Sequence Data; D009166:Mycobacterium leprae / Q000235:genetics* / Q000276:immunology; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D012641:Selection, Genetic; D017422:Sequence Analysis, DNA; D014397:Tuberculosis, Pulmonary / Q000097:blood
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D011994:Recombinant Proteins; C092234:antigen 84, Mycobacterium