title: A polymerase chain reaction-based method for the semiquantitative study of interleukin-8 mRNA in human basophil leukocytes.
abstract: We examined a potential method for quantitative analysis of cytokine expression patterns in purified human basophil leukocyte preparations. Basophil mRNA was reverse transcribed and cytokine cDNA levels determined by competitive polymerase chain reaction (PCR) with internal standard cDNAs constructed by site-directed mutagenesis. Co-reverse transcribed glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) cDNA levels were used as an internal control to correct for unequal efficiencies in RNA isolation and reverse transcription. The method was subjected to a statistical validation giving the within series precision of the analysis. This method was used to examine interleukin-8 expression patterns in basophils from different donors. The results from this study revealed increased interleukin-8 mRNA levels after in vitro challenge with a variety of stimuli.
authors.affiliation: Department of Molecular Biology, University of Aarhus, Denmark.
authors.forename: P
authors.initials: P
authors.lastname: Markvardsen
authors.forename: T
authors.initials: T
authors.lastname: Bjerke
authors.forename: N
authors.initials: N
authors.lastname: Rüdiger
authors.forename: P O
authors.initials: PO
authors.lastname: Schiøtz
authors.forename: N
authors.initials: N
authors.lastname: Gregersen
authors.forename: J
authors.initials: J
authors.lastname: Justesen
authors.forename: K
authors.initials: K
authors.lastname: Paludan
mesh_terms: D001483:Base Sequence; D001491:Basophils / Q000737:chemistry*; D018076:DNA, Complementary / Q000592:standards; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D006801:Humans; D016209:Interleukin-8 / Q000097:blood / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards; D012333:RNA, Messenger / Q000032:analysis*; D012015:Reference Standards; D013698:Templates, Genetic
chemical_list: D018076:DNA, Complementary; D016209:Interleukin-8; D012333:RNA, Messenger; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases