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Microb Pathog ; 39(3): 97-107, 2005 Sep.
Article in English | MEDLINE | ID: mdl-16098710

ABSTRACT

Identification of mycobacterial adhesins is needed to understand better the pathogenesis of tuberculosis and to develop new strategies to fight this infection. In this work, THP-1 monocytic cells were incubated with Mycobacterium tuberculosis culture filtrate proteins labelled with biotin and a dominant 19-kDa adhesin was found. This adhesin was characterized as the glycosylated and acylated 19-kDa antigen (Rv 3763). These findings were confirmed in assays with culture filtrate proteins and cell-wall fractions from a recombinant Mycobacterium smegmatis strain that overexpresses the 19-kDa antigen. Further, fluorescent microspheres coated with recombinant culture filtrate proteins adhere to cells in higher numbers than microspheres coated with native M. smegmatis proteins. The binding of the 19-kDa antigen to cells was inhibited with mannose receptor competitor sugars, Ca(2+) chelators and with a monoclonal antibody to the human mannose receptor. Phagocytosis assays showed high-level binding of bacilli to THP-1 cells that was inhibited with alpha-methyl-mannoside, mannan, EDTA and mAbs to the mannose receptor and to the 19-kDa M. tuberculosis antigen. Immunoprecipitation, cell-surface ELISA and immunostaining confirmed the expression of the mannose receptor by THP-1 cells. In conclusion, here we show that the macrophage mannose receptor, considered a pathogen pattern recognition receptor, may interact with mannose residues of mycobacterial glycoproteins that could promote the phagocytosis of mycobacteria.


Subject(s)
Adhesins, Bacterial/immunology , Lectins, C-Type/immunology , Mannose-Binding Lectins/immunology , Monocytes/immunology , Mycobacterium tuberculosis/immunology , Phagocytosis/immunology , Receptors, Cell Surface/immunology , Tuberculosis/microbiology , Acetylglucosamine/pharmacology , Adhesins, Bacterial/metabolism , Antigens, Bacterial/immunology , Bacterial Adhesion , Enzyme-Linked Immunosorbent Assay , Humans , Immunoblotting , Immunoprecipitation , Lectins, C-Type/metabolism , Mannans/pharmacology , Mannose Receptor , Mannose-Binding Lectins/metabolism , Methylmannosides/pharmacology , Monocytes/metabolism , Monocytes/microbiology , Mycobacterium tuberculosis/metabolism , Protein Binding/immunology , Receptors, Cell Surface/metabolism
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