Expression of human platelet-activating factor receptor gene in EoL-1 cells following butyrate-induced differentiation.
Platelet-activating factor (PAF) is a potent lipid mediator of allergic inflammation through its interaction with eosinophils. Expression of the PAF receptor is modulated by many agents, including those responsible for cell differentiation. We report here that differentiation of a human eosinophilic leukaemia cell line, EoL-1, by sodium n-butyrate is associated with induction of PAF receptor gene expression, as indicated by: PAF receptor mRNA accumulation; increases in the binding of [3H]WEB 2086, a PAF antagonist; analysis of cell-surface expression of PAF receptor protein using a monoclonal anti-(PAF receptor) antibody; and augmentation of PAF-induced increase in the intracellular concentration of calcium. Using cDNA cloning, the receptor expressed in EoL-1 cells was identified as 'Transcript 1', one of two transcripts which was previously reported from human genomic analysis (Mutoh, Bito, Minami, Nakamura, Honda, Izumi, Nakata, Kurachi, Terano and Shimizu (1993) FEBS Lett. 322, 129-134). The PAF-induced calcium response and phosphoinositide turnover were decreased by pertussis toxin (PTX) treatment, suggesting that these signals are coupled largely with PTX-sensitive G-protein(s) in EoL-1 cells. These systems may provide a useful experimental model with which to investigate the relationship between eosinophilic differentiation and PAF receptor induction, and the role of eosinophils in allergic responses.
Department of Biochemistry, Faculty of Medicine, University of Tokyo, Japan.
T
T
Izumi
S
S
Kishimoto
T
T
Takano
M
M
Nakamura
Y
Y
Miyabe
M
M
Nakata
C
C
Sakanaka
T
T
Shimizu
D000595:Amino Acid Sequence; D000818:Animals; D001381:Azepines / Q000378:metabolism; D001483:Base Sequence; D002087:Butyrates / Q000494:pharmacology*; D020148:Butyric Acid; D016466:CHO Cells; D002454:Cell Differentiation / Q000187:drug effects*; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D015870:Gene Expression; D006168:Guinea Pigs; D006801:Humans; D017681:Hypereosinophilic Syndrome / Q000378:metabolism* / Q000473:pathology; D015544:Inositol 1,4,5-Trisphosphate / Q000096:biosynthesis; D008969:Molecular Sequence Data; D037342:Pertussis Toxin; D010980:Platelet Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D017422:Sequence Analysis, DNA; D014230:Triazoles / Q000378:metabolism; D014316:Tritium; D014407:Tumor Cells, Cultured; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D001381:Azepines; D002087:Butyrates; D018076:DNA, Complementary; D010980:Platelet Membrane Glycoproteins; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D014230:Triazoles; D010566:Virulence Factors, Bordetella; C039351:platelet activating factor receptor; D014316:Tritium; C052518:WEB 2086; D020148:Butyric Acid; D015544:Inositol 1,4,5-Trisphosphate; D037342:Pertussis Toxin