Quantitative evaluation of macrophage phagocytosing capacity by a fluorometric assay.
This paper reviews sensitive and simple quantitative evaluation of macrophage phagocytosing capacity by applying fluoresecin-labeled Sacharomyces cerevisiae cells. Yeast cells were conjugated with fluoresceinisothiocyanate (FITC) and used as fluorescent particles. A time course analysis within this method showed that phagocytosis of yeast cells was temperature dependent and that the number of that ones ingested by macrophages increased rapidly during the initial 60 min of incubation at 37 degrees C. Free fluorescent cells can be effectively removed by aspiration from the well. Furthermore, yeast cells required preopsonization with serum to achieve optimal uptake of the cells. The uptake of nonopsonized yeast cells by macrophages was significantly lower than that of opsonized cells (P < 0.05). We propose that about 50% of mouse macrophages can carry functionally active FcR responsible for phagocytosis.
Miliukiene
Vale
V
Department of Developmental Biology, Institute of Biochemistry, 12 Mokslininku Street, LT-2600, Vilnius, Lithuania. aiste@ifo.it
Biziuleviciene
Gene
G
Pilinkiene
Audrone
A
D000818:Animals; D001681:Biological Assay / Q000379:methods*; D005260:Female; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D017737:Macrophages, Peritoneal / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010587:Phagocytosis / Q000502:physiology*; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000378:metabolism; D013696:Temperature; D013997:Time Factors
D005456:Fluorescent Dyes; D016650:Fluorescein-5-isothiocyanate