title: The production and characterization of a modified recombinant interleukin-1 receptor antagonist.
abstract: The increasingly complex cytokine network involves both positive and negative regulatory pathways. Natural inhibitors of cytokines are of great importance both as analytical tools and as potential therapeutic agents. Interleukin-1 (IL-1) inhibitory bioactivity including a specific receptor antagonist of IL-I (IL-1ra) has been described both in cultured cell supernatants and in human body fluids. In the current studies, the cDNA of IL-1ra from human monocytes was obtained by the techniques of mRNA isolation and reverse transcription/polymerase chain reaction (RT/PCR). The IL-1ra cDNA/pET-15b was transfected into DE3 cells and the recombinant protein expressed. The purified protein was demonstrated as a single band with molecular mass of 20 KD by SDS-PAGE; it had strong IL-1 inhibitory activity. This IL-1 inhibitor competed with IL-1 for their receptor as assessed by flow cytometer. The existence of this naturally occurring specific cytokine receptor antagonist may lead to a different perspective of the cytokine network. The availability of this recombinant IL-1 receptor antagonist allows us to test its role on the cytokine network, and on possible disease modification.
authors.affiliation: Tri-Service General Hospital, Taipei, Taiwan, R.O.C.
authors.forename: D M
authors.initials: DM
authors.lastname: Chang
authors.forename: W Y
authors.initials: WY
authors.lastname: Chou
mesh_terms: D001483:Base Sequence; D018076:DNA, Complementary / Q000302:isolation & purification; D006801:Humans; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1 / Q000037:antagonists & inhibitors; D008969:Molecular Sequence Data; D017472:Receptors, Interleukin-1 / Q000037:antagonists & inhibitors*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D012795:Sialoglycoproteins / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D018076:DNA, Complementary; C507763:IL1RN protein, human; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1; D017472:Receptors, Interleukin-1; D011994:Recombinant Proteins; D012795:Sialoglycoproteins