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.
N1 - Funding Information:
This work was supported by the American Lung Association of Nebraska and by the James M. Keck Faculty Development Award from the Health Future Foundation to D.K.A. We would like to thank Dr. J. Rokach for synthetic LTs and Dr. R. Hughes for BC3H-1 cells. We also thank Mrs. Rosemarv Batts for typing the manuscript.
Copyright:
Copyright 2018 Elsevier B.V., All rights reserved.
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.
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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 -