Association of a macrophage galactoside-binding protein with Mycobacterium-containing phagosomes.
Mycobacteria reside intracellularly in a vacuole that allows it to circumvent the antimicrobial environment of the host macrophage. Although the mycobacterial phagosome exhibits selective fusion with vesicles of the endosomal system, identification of host and bacterial factors associated with phagosome bio-genesis is limited. To identify these potential factors, mAbs were generated to a membrane preparation of mycobacterial phagosomes isolated from M. tuberculosis-infected macrophages. A mAb recognizing a 32-35 kDa macrophage protein associated with the phagosomal membrane of Mycobacterium was identified. N-terminal sequence analysis identified this protein as Mac-2 or galectin-3, a galactoside-binding protein of macrophages. Galectin-3 (gal-3) was shown to accumulate in Mycobacterium-containing phagosomes during the course of infection. This accumulation was specific for phagosomes containing live mycobacteria and occurred primarily at the cytosolic face of the phagosome membrane. In addition, bind-ing of gal-3 to mycobacterial phosphatidylinositol mannosides (PIMs) demonstrated a novel interaction between host carbohydrate-binding proteins and released mycobacterial glycolipids. Infection of macrophages from gal-3-deficient mice indicated that the protein did not play a role in infection in vitro. In contrast, infection of gal-3-deficient mice revealed a reduced capacity to clear late but not early infection.
Beatty
Wandy L
WL
Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO 63110, USA. beatty@borcim.wustl,com
Rhoades
Elizabeth R
ER
Hsu
Daniel K
DK
Liu
Fu-Tong
FT
Russell
David G
DG
D000818:Animals; D000943:Antigens, Differentiation / Q000235:genetics / Q000378:metabolism*; D002417:Cattle; D005697:Galactosides / Q000378:metabolism; D037502:Galectin 3; D006801:Humans; D008264:Macrophages / Q000378:metabolism / Q000382:microbiology; D051379:Mice; D008807:Mice, Inbred BALB C; D009163:Mycobacterium bovis / Q000254:growth & development / Q000472:pathogenicity; D009169:Mycobacterium tuberculosis / Q000254:growth & development / Q000472:pathogenicity*; D010588:Phagosomes / Q000382:microbiology*
D000943:Antigens, Differentiation; D005697:Galactosides; D037502:Galectin 3