Stimulation of leukotriene production and membrane translocation of 5-lipoxygenase by cross-linking of the IgE receptors in RBL-2H3 cells.
Recent studies in rat basophilic leukemia cells (RBL-2H3) have shown that two pharmacological agents, ionomycin and thapsigargin, induce leukotriene C4 production and translocation of 5-lipoxygenase from cytosol to membrane, primarily by causing an influx of extracellular calcium. In the present study, we investigate the induction of these events by receptor activation. Cross-linking of high-affinity IgE receptors (Fc epsilon RI) by antigen in RBL-2H3 cells leads to leukotriene C4 production and membrane translocation of 5-lipoxygenase. As in the ionomycin-stimulated cells, leukotriene C4 production in antigen-stimulated cells is calcium-dependent since the amount of leukotriene C4 produced correlates quantitatively with the increase in intracellular free calcium concentration ([Ca2+]i). However, the increase in [Ca2+]i required for equivalent leukotriene C4 production by antigen is not as high as it is using ionomycin. In addition, no threshold [Ca2+]i level is required for leukotriene production by antigen, which is in contrast to the ionomycin stimulation that a [Ca2+]i level of 300-400 nM is required. Furthermore, antigen causes an additive increase in leukotriene C4 production in cells stimulated by the ionomycin. These results suggest that another as yet unidentified intracellular pathway acts in conjunction with Ca2+ for leukotriene synthesis in antigen-stimulated cells. Antigen stimulation causes 20-30% of the total cell 5-lipoxygenase to associate with membranes (compared with 10% in unstimulated cells) as demonstrated by enzyme activity assay and by Western Blot using antibodies to 5-lipoxygenase.(ABSTRACT TRUNCATED AT 250 WORDS)
Wong
A
A
Department of Cellular Biochemistry and Immunology, SmithKline Beecham Pharmaceuticals, King of Prussia, Pennsylvania 19406.
Cook
M N
MN
Hwang
S M
SM
Sarau
H M
HM
Foley
J J
JJ
Crooke
S T
ST
D000818:Animals; D000941:Antigens / Q000276:immunology; D000944:Antigens, Differentiation, B-Lymphocyte / Q000276:immunology*; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism*; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology; D002462:Cell Membrane / Q000201:enzymology; D003432:Cross-Linking Reagents; D007211:Indoles / Q000494:pharmacology; D015759:Ionomycin / Q000494:pharmacology; D015471:Leukemia, Basophilic, Acute / Q000276:immunology* / Q000378:metabolism; D051381:Rats; D011961:Receptors, Fc / Q000276:immunology*; D017455:Receptors, IgE; D013189:SRS-A / Q000096:biosynthesis*; D014407:Tumor Cells, Cultured; D002722:Chlorobenzenes
D000941:Antigens; D000944:Antigens, Differentiation, B-Lymphocyte; D001094:Arachidonate 5-Lipoxygenase; D002118:Calcium; D003432:Cross-Linking Reagents; D007211:Indoles; D015759:Ionomycin; D011961:Receptors, Fc; D017455:Receptors, IgE; D013189:SRS-A; C060893:MK-886; D002722:Chlorobenzenes