Flow cytometric assay of lung macrophage uptake of environmental particulates
- 1 May 1995
- Vol. 20 (1), 23-32
- https://doi.org/10.1002/cyto.990200106
Abstract
We sought to establish a quantitative method using flow cytometry to study uptake of environmental particulates by alveolar macrophages (AMs). We used right angle light scatter (RAS) to measure uptake of titanium dioxide, quartz, and diesel particulates. After incubation with TiO2 in vitro, AMs showed dose‐dependent increases in both cell‐associated particles visualized by microscopy and RAS measured by flow cytometry (e.g., fold increase RAS at 4, 8, 16, 32, and 80 μ/ml, respectively, = 2 ± 0.1, 4.0 ± 0.5, 5.5 ± 0.5, 9.1 ± 2.5, 14.3 ± 0.9; mean ± SEM). Similar results were obtained with quartz and diesel particles. A strong correlation was observed between particle load per cell and AM RAS after uptake of fluorescent latex beads or fluorescent TiO2 (coated with BODIPY‐BSA) (R2 = 0.984, 0.997, respectively). Using this technique, we found AM uptake of environmental particulates to be substantially greater than that of a panel of myelomonocytic and epithelial cell lines, consistent with their physiologic role in pulmonary defenses. RAS measurements have also identified both calcium‐dependent and calcium‐independent components in AM interactions with inert particles. Although this technique does not allow precise quantitation of particle number or mass per cell, flow cytometric analysis of relative increases in RAS is a useful tool to study AM interactions with a variety of environmental particulates.Keywords
This publication has 17 references indexed in Scilit:
- What Are People Dying of on High Air Pollution Days?Environmental Research, 1994
- Air Pollution and Daily Mortality: A Review and Meta AnalysisEnvironmental Research, 1994
- An Association between Air Pollution and Mortality in Six U.S. CitiesNew England Journal of Medicine, 1993
- A fluorescence technique to distinguish attached from ingested erythrocytes and zymosan particles in phagocytosing macrophagesJournal of Immunological Methods, 1991
- The use of fluorescence quenching in flow cytofluorometry to measure the attachment and ingestion phases in phagocytosis in peripheral blood without prior cell separationJournal of Immunological Methods, 1987
- Macrophages, Dust, and Pulmonary DiseasesExperimental Lung Research, 1987
- Quantitative evaluation of opsonin-independent phagocytosis by alveolar macrophages in monolayer using polystyrene microspheresJournal of Immunological Methods, 1985
- Flow cytometric assay for combined measurement of phagocytosis and intracellular killing of Candida albicansJournal of Immunological Methods, 1984
- An in vivo hamster bioassay to assess the toxicity of particulates for the lungsToxicology and Applied Pharmacology, 1982
- Effects of a Brief Low-Level Exposure to the Particulate Fraction of Diesel Exhaust on Pulmonary Function of Conscious SheepArchives of environmental health, 1980