Immunol Cell Biol. 1996 Aug;74(4):306-12. Unique Identifier : AIDSLINE
Studies of phagocytic efficiency in cells of the macrophage lineage have
assumed additional importance since the discovery that HIV infection of
these cells impairs their immune function. A rapid method has been
developed for measuring phagocytosis of the opportunistic pathogen
Mycobacterium avium complex by human monocytes. Fluoresceinated M. avium
complex (F-MAC) was incubated with whole blood at 37 degrees C and the
fluorescence of extracellular F-MAC was quenched using a vital blue
stain. Monocytes were then stained with a monoclonal antibody (mAb) to
human CD14 conjugated to phycoerythrin (PE) red cells were lysed, and
the percentage of monocytes which had phagocytosed F-MAC was measured by
flow cytometry. The results were reproducible in samples of blood taken
from individual donors over a period of 1 or 2 weeks, and optimum F-MAC
concentrations and an optimum incubation time were determined by
experiment. This method has the advantages of requiring only a small
volume of blood, not necessitating manipulation of cells before testing,
and using a phagocytic target relevant to the pathogenesis of HIV
infection.
Fluorescein-5-isothiocyanate/CHEMISTRY Human Monocytes/*IMMUNOLOGY
Mycobacterium avium Complex/*IMMUNOLOGY Phagocytosis/*IMMUNOLOGY
Reproducibility of Results Support, Non-U.S. Gov't JOURNAL ARTICLE