A specific radioreceptor assay for leukotriene C4 and the measurement of calcium ionophore-induced leukotriene C4 production from human leukocytes

Tamura Naohiko, Devendra K. Agrawal, Robert G. Townley

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Pages (from-to)327-333
Number of pages7
JournalJournal of Pharmacological Methods
Volume18
Issue number4
DOIs
StatePublished - 1987

Fingerprint

Leukotriene C4
Radioligand Assay
Calcium Ionophores
Assays
Leukocytes
Eosinophils
Calcimycin
Cell membranes
Neutrophils
Cell Membrane
Leukotriene E4
Leukotriene D4
Inhibitory Concentration 50
Recovery

All Science Journal Classification (ASJC) codes

  • Pharmacology

Cite this

@article{42588112c1e3411bbe938d1372af1cf1,
title = "A specific radioreceptor assay for leukotriene C4 and the measurement of calcium ionophore-induced leukotriene C4 production from human leukocytes",
abstract = "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.",
author = "Tamura Naohiko and Agrawal, {Devendra K.} and Townley, {Robert G.}",
year = "1987",
doi = "10.1016/0160-5402(87)90064-7",
language = "English",
volume = "18",
pages = "327--333",
journal = "Journal of Pharmacological and Toxicological Methods",
issn = "1056-8719",
publisher = "Elsevier Inc.",
number = "4",

}

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

Y1 - 1987

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

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 -