Aerobic responses of mouse macrophages to phagocytosis against Escherichia coli as revealed by microphotometry.
Fluorescence of peritoneal macrophages, which ingested E. coli labelled with N-[7-(dimethylamino)-4-methyl-coumarinyl]-maleimide, was measured with a microfluorometer. The cellular contents of protein and formazan of a monotetrazolium produced by succinate dehydrogenase (SD) activity were assayed with a microspectrophotometer. During phagocytosis, protein increased in 30 min., but was balanced later by self-hydrolysis. SD activity was unchanged for 30 min. and then increased until 2 hrs. Upon removal of bacteria, the fluorescence and protein decreased with time, whereas SD activity increased further for 1 hr. before decrease. This indicates a delayed activation of the enzyme. The cellular ATP content was reduced during phagocytosis and restored to the normal level 3 hrs. after removal of bacteria. Both KCN-sensitive and -insensitive oxygen uptakes increased in phagocytosis. Thus, oxidative metabolism of macrophages is stimulated in phagocytosis and the stimulation can be demonstrated in situ by the microphotometric methods.
Shono
M
M
General Laboratories for Medical Research, School of Medicine, University of Tokushima, Japan.
Yamada
M
M
Yamaguchi
H
H
Miyamoto
H
H
D000255:Adenosine Triphosphate / Q000378:metabolism; D000332:Aerobiosis; D000818:Animals; D004926:Escherichia coli; D005470:Fluorometry; D008264:Macrophages / Q000502:physiology*; D008297:Male; D051379:Mice; D008817:Mice, Mutant Strains; D010101:Oxygen Consumption / Q000187:drug effects; D010587:Phagocytosis; D011190:Potassium Cyanide / Q000494:pharmacology
D000255:Adenosine Triphosphate; D011190:Potassium Cyanide