Isolation of functionally active pleural macrophages.
Pleural fluid provides an easily accessible source of tissue macrophages (MACS). We established a method for retrieval and isolation of these cells. Thirty thoracenteses were performed on patients with the clinical diagnosis of congestive heart failure or malignancy. Within 4 hr of thoracentesis, the specimen was placed on a Ficoll-Hypaque gradient, centrifuged, and the mononuclear cells were isolated. Assessment for total cell count, cell viability, and percentage of cells which were MACS as determined by nonspecific esterase staining was recorded. Pleural MACS were separated by glass adherence. In 24 cases, sufficient cells were available to measure the release of hydrogen peroxide spontaneously and with phorbol myristate acetate (PMA) stimulation. Nine of 24 specimens had spontaneous hydrogen peroxide release. In all cases, stimulation of pleural MACS resulted in a significant increase in the amount of hydrogen peroxide released. We report a simple method for retrieving viable and functionally active pleural macrophages.
Baughman
R P
RP
Department of Medicine, University of Cincinnati Medical Center, Ohio.
Doers
J
J
Lower
E E
EE
Buchsbaum
J
J
D002452:Cell Count; D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D006333:Heart Failure / Q000150:complications / Q000473:pathology; D006801:Humans; D006861:Hydrogen Peroxide / Q000032:analysis; D008262:Macrophage Activation / Q000187:drug effects; D008264:Macrophages / Q000378:metabolism / Q000502:physiology; D009369:Neoplasms / Q000150:complications / Q000473:pathology; D010996:Pleural Effusion / Q000209:etiology / Q000473:pathology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D006861:Hydrogen Peroxide; D013755:Tetradecanoylphorbol Acetate