Phagocytosis of a fluorescently labeled perflubron emulsion by a human monocyte cell line.
We hypothesized that fluorocarbon-based lipid emulsions are phagocytosed by monocytes and that many of the in vivo side effects related to the infusion of these particulate emulsions are due to release of cytokines by these monocytes. To clarify whether these emulsions are actually phagocytosed we attempted to measure by flow cytometry the apparent uptake of a fluorescently labeled high-concentration (90%, w/v) perflubron (perfluorooctyl bromide [PFOB]) emulsion by a differentiated human monocyte cell line. A fluorescent chromophore (Zynaxis Cell Science) was used to label the egg yolk phospholipid in a perflubron emulsion. This phospholipid label was used to track the perflubron emulsion during overnight incubation with the human monocyte (THP-1) cell line which had been differentiated, by exposure to PMA, into macrophage-like cells. Our results indicate that after 24 hours of incubation with the labeled perflubron emulsion, 64.9% (+/- 11.0) of differentiated THP-1 cells had cell-associated emulsion (ingested and/or membrane bound) whereas 24.4 (+/- 6.8%) of the control cells had cell-associated emulsion. We speculate that this technique may be a useful method to track the intravascular persistence and extravascular distribution of such emulsions, and that the degree of uptake of the emulsion by macrophages in this assay may correlate with its in vivo half life.
Alliance Pharmaceutical Corporation, San Diego, CA 92121.
D J
DJ
Smith
E S
ES
Kornbrust
T A
TA
Lane
D002460:Cell Line; D003571:Cytochalasin B; D004655:Emulsions; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D005466:Fluorocarbons / Q000097:blood*; D006801:Humans; D006842:Hydrocarbons, Brominated; D009000:Monocytes / Q000502:physiology*; D010587:Phagocytosis / Q000502:physiology*
D004655:Emulsions; D005456:Fluorescent Dyes; D005466:Fluorocarbons; D006842:Hydrocarbons, Brominated; D003571:Cytochalasin B; C003072:perflubron