title: [Application of NF-kappaB reporter and Dual-Luciferase assays in the measure of bioactivity of interleukin-1beta and interleukin-1 receptor antagonist].
abstract: OBJECTIVE
  To develop a reporter gene system based on transient transfections with a NF-kappaB responsive reporter gene to detect the bioactivity of IL-1beta and IL-1 receptor antagonist.
  
  
  METHODS
  NF-kappaB reporter and Dual-Luciferase assays were applied to measure the bioactivity of IL-1beta and IL-1 receptor antagonist in mouse EL4 cells (some subclones of EL4 cells expressed high level of IL-1 receptor on cell surface). pNF-kappaB-luc and pRL-TK, used as an internal control, were co-transfected into EL4 cells and then the IL-1beta was added.
  
  
  RESULTS
  The results indicated that IL-1beta was able to induce the expression of this luciferase, which could be blocked by IL-1 receptor antagonist. The optimal dose of IL-1beta was 5 microg/L in Dual-Luciferase assay, whose bioactivity can be effectively inhibited by IL-1ra at 50 microg/L.
  
  
  CONCLUSIONS
  We have established a new method to detect the bioactivity of IL-1beta and IL-1 receptor antagonist, which can give repeatable results.
authors.affiliation: Department of Immunology, Peking University School of Basic Medical Science, Peking University, Beijing 100083, China.
authors.forename: Ying-mei
authors.initials: YM
authors.lastname: Zhang
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.forename: Qian-mei
authors.initials: QM
authors.lastname: Xu
authors.forename: Bing-feng
authors.initials: BF
authors.lastname: Lv
authors.forename: Da-long
authors.initials: DL
authors.lastname: Ma
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D053590:Interleukin 1 Receptor Antagonist Protein / Q000494:pharmacology*; D053583:Interleukin-1beta / Q000494:pharmacology*; D008156:Luciferases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D017472:Receptors, Interleukin-1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D013945:Thymoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D013953:Thymus Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D014162:Transfection
chemical_list: D053590:Interleukin 1 Receptor Antagonist Protein; D053583:Interleukin-1beta; D016328:NF-kappa B; D017472:Receptors, Interleukin-1; D011993:Recombinant Fusion Proteins; D008156:Luciferases