title: BODIPY(®) FL histamine as a new modality for quantitative detection of histamine receptor upregulation upon IgE sensitization in murine bone marrow-derived mast cells.
abstract: Flow cytometry is one of the most widely used methods for the qualitative and quantitative analysis of cell surface expressed proteins by making use of fluorescent specific antibodies. Lacking an antibody validated for flow cytometry, an alternative approach for labeling cell surface receptors is the use of fluorescently tagged ligands. In this study, histamine H4 receptor transfected Chinese hamster ovary cells and murine bone marrow-derived mast cells (mBMMCs) were selected for studying the possibility of staining individual histamine receptors using BODIPY(®) FL histamine and selective antagonists. Flow cytometric measurements and supporting calculations showed that BODIPY FL histamine is suitable tool for quantitating cell surface histamine receptors. The binding, and competitive inhibition of this fluorescent ligand were characterized, which were in good agreement with a semi-empirical model constructed from fundamental protein-binding relationships. Using this method it was shown for the first time that even though mature mBMMCs express H2R and H4R to the same extent, immunoglobulin E sensitization results in H4R upregulation only, while the surface expression of H2R remains unchanged.
authors.affiliation: Sanofi Research and Development, Tó utca 1-5, H-1045, Budapest, Hungary; Faculty of Pharmacy, Semmelweis University, Hőgyes Endre utca 9, H-1092, Budapest, Hungary.
authors.forename: Arash
authors.initials: A
authors.lastname: Mirzahosseini
authors.forename: Marianna
authors.initials: M
authors.lastname: Kovács
authors.forename: Károly
authors.initials: K
authors.lastname: Kánai
authors.forename: Péter
authors.initials: P
authors.lastname: Csutora
authors.forename: Balázs
authors.initials: B
authors.lastname: Dalmadi
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D001896:Boron Compounds / Q000737:chemistry; D016466:CHO Cells; D003412:Cricetulus; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D015870:Gene Expression; D006632:Histamine / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007073:Immunoglobulin E / Q000494:pharmacology; D008297:Male; D008407:Mast Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D061251:Primary Cell Culture; D011485:Protein Binding; D043562:Receptors, G-Protein-Coupled / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D011968:Receptors, Histamine / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D011970:Receptors, Histamine H2 / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D000074040:Receptors, Histamine H4; D014162:Transfection
chemical_list: C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D005456:Fluorescent Dyes; C418607:HRH4 protein, human; D043562:Receptors, G-Protein-Coupled; D011968:Receptors, Histamine; D011970:Receptors, Histamine H2; D000074040:Receptors, Histamine H4; D007073:Immunoglobulin E; D006632:Histamine
keywords: IgE sensitization; flow cytometry; fluorescent ligand; histamine H4 receptor; mast cell