The 19-kDa antigen of Mycobacterium tuberculosis is a major adhesin that binds the mannose receptor of THP-1 monocytic cells and promotes phagocytosis of mycobacteria.
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.
Diaz-Silvestre
Hugo
H
Departamento de Inmunologia, Instituto de Investigaciones Biomedicas, Universidad Nacional Autonoma de Mexico, Apartado Postal 70-228, 04510 Mexico, DF, Mexico.
Espinosa-Cueto
Patricia
P
Sanchez-Gonzalez
Alejandro
A
Esparza-Ceron
Miguel A
MA
Pereira-Suarez
Ana Laura
AL
Bernal-Fernandez
German
G
Espitia
Clara
C
Mancilla
Raul
R
D000117:Acetylglucosamine / Q000494:pharmacology; D018829:Adhesins, Bacterial / Q000276:immunology* / Q000378:metabolism; D000942:Antigens, Bacterial / Q000276:immunology; D001422:Bacterial Adhesion; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D037181:Lectins, C-Type / Q000276:immunology* / Q000378:metabolism; D008351:Mannans / Q000494:pharmacology; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins / Q000276:immunology* / Q000378:metabolism; D008766:Methylmannosides / Q000494:pharmacology; D009000:Monocytes / Q000276:immunology* / Q000378:metabolism / Q000382:microbiology; D009169:Mycobacterium tuberculosis / Q000276:immunology* / Q000378:metabolism; D010587:Phagocytosis / Q000276:immunology*; D011485:Protein Binding / Q000276:immunology; D011956:Receptors, Cell Surface / Q000276:immunology* / Q000378:metabolism; D014376:Tuberculosis / Q000382:microbiology*
D018829:Adhesins, Bacterial; D000942:Antigens, Bacterial; D037181:Lectins, C-Type; D008351:Mannans; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D008766:Methylmannosides; D011956:Receptors, Cell Surface; C008466:methylmannoside; D000117:Acetylglucosamine