title: Mycobacterium leprae produces extracellular homologs of the antigen 85 complex.
abstract: The antigen 85 complex is a set of at least three closely related secreted proteins (85A, 85B, and 85C) of 30 to 32 kDa produced by Mycobacterium tuberculosis and other mycobacteria. Their prominence in Mycobacterium leprae, the one obligate intracellular pathogen of the genus, had been assumed on the basis of immunological evidence and proof of the existence of the gene encoding the 85B protein of the complex. We have now observed the production of this family of proteins by M. leprae through analysis of various fractions by Western blotting (immunoblotting) with monospecific rabbit antisera raised against the individual Mycobacterium bovis BCG 85A, 85B, and 85C proteins. A predominant cross-reactive band with an apparent molecular mass of 30 kDa was detected in extracts of nondisrupted whole M. leprae and in soluble fractions prepared from the tissues of M. leprae-infected armadillos. Further studies of the subcellular distribution of this protein within the bacterium confirmed that it is secreted by the organism, an observation that explains past difficulties in detecting the antigen 85 complex in M. leprae. Confirmation that the M. leprae product is a member of the antigen 85 complex was obtained by comparison of peptide fingerprints with those from the BCG product. The pattern of reactivity of the M. leprae antigen 85 complex with anti-M. bovis BCG 85B serum, as well as two-dimensional electrophoresis, established that the 85B component was the predominant member of the complex in M. leprae. The fibronectin-binding capacity of the M. leprae and BCG 85 complexes was reinvestigated by new approaches and is questioned. Nevertheless, the results obtained with the native proteins reinforce previous reports, derived primarily from the use of homologous proteins, that the antigen 85 complex is one of the dominant protein immunogens of the leprosy bacillus.
authors.affiliation: Department of Microbiology, Colorado State University, Fort Collins 80523.
authors.forename: M C
authors.initials: MC
authors.lastname: Pessolani
authors.forename: P J
authors.initials: PJ
authors.lastname: Brennan
mesh_terms: D000942:Antigens, Bacterial / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000276:immunology; D003429:Cross Reactions; D015180:Electrophoresis, Gel, Two-Dimensional; D005110:Extracellular Space / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D008970:Molecular Weight; D009163:Mycobacterium bovis / Q000276:immunology; D009166:Mycobacterium leprae / Q000276:immunology* / Q000648:ultrastructure; D011485:Protein Binding
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D005353:Fibronectins; C071448:antigen 85B, Mycobacterium leprae