TY - JOUR
T1 - A specific radioreceptor assay for leukotriene C4 and the measurement of calcium ionophore-induced leukotriene C4 production from human leukocytes
AU - Naohiko, Tamura
AU - Agrawal, Devendra K.
AU - Townley, Robert G.
PY - 1987/12
Y1 - 1987/12
N2 - A highly specific radioreceptor assay (RRA) for leukotriene (LT) C4 using BC3H-1 cell membrane as a source of LTC4 receptors was developed and demonstrated its use in the measurement of calcium ionophore A23187-induced LTC4 production from purified eosinophils, neutrophils, and mononuclear cells. Unlabeled LTC4 competed for the specific [3H]LTC4 binding to the BC3H-1 cell membranes in a dose-dependent manner. Under the experimental conditions used in this study, the calculated IC50 value of unlabeled LTC4 was 27.2 ± 1.2 nM (n = 5). The sensitivity of the method for LTC4 was 0.6 pmol. The cross-reactivities of LTD4, LTE4, and FPL 55712 were negligible. The recovery of the exogenously added LTC4 with eosinophils was greater than 90% in both the presence and the absence of calcium ionophore A23187. Calcium ionophore induced 73.0 ± 17.0 ng of LTC4 production/106 eosinophils (n = 8), and this was about 20-40 times more than those from neutrophils and from mononuclear cells.
AB - A highly specific radioreceptor assay (RRA) for leukotriene (LT) C4 using BC3H-1 cell membrane as a source of LTC4 receptors was developed and demonstrated its use in the measurement of calcium ionophore A23187-induced LTC4 production from purified eosinophils, neutrophils, and mononuclear cells. Unlabeled LTC4 competed for the specific [3H]LTC4 binding to the BC3H-1 cell membranes in a dose-dependent manner. Under the experimental conditions used in this study, the calculated IC50 value of unlabeled LTC4 was 27.2 ± 1.2 nM (n = 5). The sensitivity of the method for LTC4 was 0.6 pmol. The cross-reactivities of LTD4, LTE4, and FPL 55712 were negligible. The recovery of the exogenously added LTC4 with eosinophils was greater than 90% in both the presence and the absence of calcium ionophore A23187. Calcium ionophore induced 73.0 ± 17.0 ng of LTC4 production/106 eosinophils (n = 8), and this was about 20-40 times more than those from neutrophils and from mononuclear cells.
UR - http://www.scopus.com/inward/record.url?scp=0023505292&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0023505292&partnerID=8YFLogxK
U2 - 10.1016/0160-5402(87)90064-7
DO - 10.1016/0160-5402(87)90064-7
M3 - Article
C2 - 3695541
AN - SCOPUS:0023505292
VL - 18
SP - 327
EP - 333
JO - Journal of Pharmacological and Toxicological Methods
JF - Journal of Pharmacological and Toxicological Methods
SN - 1056-8719
IS - 4
ER -