Applying label-free dynamic mass redistribution assay for studying endogenous FPR1 receptor signalling in human neutrophils.
J Pharmacol Toxicol Methods
; 88(Pt 1): 72-78, 2017 Nov.
Article
em En
| MEDLINE
| ID: mdl-28716665
ABSTRACT
INTRODUCTION:
The label-free dynamic mass redistribution-based assay (DMR) is a powerful method for studying signalling pathways of G protein-coupled receptors (GPCRs). Herein we present the label-free DMR assay as a robust readout for pharmacological characterization of formyl peptide receptors (FPRs) in human neutrophils.METHODS:
Neutrophils were isolated from fresh human blood and their responses to FPR1 and FPR2 agonists, i.e. compound 43, fMLF and WKYMVm were measured in a label-free DMR assay using Epic Benchtop System from Corning®. Obtained DMR traces were used to calculate agonist potencies.RESULTS:
The potencies (pEC50) of fMLF, WKYMVm and compound 43, determined on human neutrophils using the label-free DMR assay were 8.63, 7.76 and 5.92, respectively. The DMR response to fMLF, but not WKYMVm and compound 43 could be blocked by the FPR1-specific antagonist cyclosporin H.DISCUSSION:
We conclude that the DMR assay can be used, and complements more traditional methods, to study the signalling and pharmacology of endogenous FPR receptors in human neutrophils.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Bioensaio
/
Técnicas Biossensoriais
/
Transdução de Sinais
/
Receptores de Formil Peptídeo
/
Neutrófilos
Limite:
Humans
Idioma:
En
Revista:
J Pharmacol Toxicol Methods
Assunto da revista:
FARMACOLOGIA
/
TOXICOLOGIA
Ano de publicação:
2017
Tipo de documento:
Article