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

Microb Pathog. 2005 Sep;39(3):97-107. doi: 10.1016/j.micpath.2005.06.002.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

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

Substances

  • Adhesins, Bacterial
  • Antigens, Bacterial
  • Lectins, C-Type
  • Mannans
  • Mannose Receptor
  • Mannose-Binding Lectins
  • Methylmannosides
  • Receptors, Cell Surface
  • methylmannoside
  • Acetylglucosamine