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Quantifying phagocytosis of Mycobacterium avium complex by human monocytes in whole blood.
Hewish MJ; Meikle AM; Hunter SD; Crowe SM; AIDS Pathogenesis Research
March 30, 1997
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