title: Inhibition of interleukin-1 beta mRNA expression and interleukin-1 alpha and beta secretion by a specific human recombinant interleukin-1 receptor antagonist in human peripheral blood mononuclear cells.
abstract: The transcription and translation of interleukin-1 (IL-1) may have a pleiotropic effect on the immune system and inflammatory diseases. Recently it has been reported that human monocytes not only produce IL-1 but also induce, with adherent IgG, the secretion of an IL-1 receptor antagonist (IL-1Ra), which can play an essential in vivo and in vitro role in the regulation of IL-1 activity. Recombinant human (rh) IL-1Ra is structurally similar to IL-1 beta but with no IL-1-like activity, and specifically binds to the IL-1 receptor. To more fully evaluate and clarify the inhibitory effect of rhIL-1 receptor antagonist on IL-1 we have studied the influence of rhIL-1Ra on IL-1 transcription and translation. In this report we show that IL-1 beta mRNA from peripheral blood mononuclear cells (PBMC) is strongly inhibited (66%) when rhIL-1Ra (250 ng/ml) was added to cultured cells activated with lipopolysaccharide (LPS) (100 ng/ml) for 4 hr, determined with the slot blot analysis. The addition of exogenous rhIL-1 beta to the cell culture diminished the inhibitory effect (44%). Moreover, we report that the block of IL-1 mRNA transcription consequently leads to the inhibition of IL-1 alpha and IL-1 beta secretion in human PBMC, as measured by ELISA method. In fact, herein we show that LPS activates human PBMC to secrete IL-1 beta and IL-1 alpha, an effect inhibited, in a dose-dependent fashion by rhIL-1Ra (0.025-250 ng/ml) in an overnight incubation. Since IL-1 is a strong inducer of IL-1 synthesis in vivo and in vitro, in our study we used rh IL-1 alpha to stimulate the secretion of IL-1 beta in human PBMC. This activation, carried out overnight, also provoked the release of IL-1 beta in a dose-dependent manner, which was strongly inhibited by rhIL-1Ra used at different concentrations (0.025-250 ng/ml). The inhibitory effect exerted by IL-1Ra on human PBMC IL-1 mRNA transcription and the down-regulation of secretion of IL-1 beta stimulated by IL-1 alpha, may contribute to therapeutic effects in inflammatory diseases such as rheumatoid arthritis and other autoimmune diseases.(ABSTRACT TRUNCATED AT 400 WORDS)
authors.affiliation: Immunology Division, University of Chieti Medical School, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Conti
authors.forename: C
authors.initials: C
authors.lastname: Feliciani
authors.forename: R C
authors.initials: RC
authors.lastname: Barbacane
authors.forename: M R
authors.initials: MR
authors.lastname: Panara
authors.forename: M
authors.initials: M
authors.lastname: Reale
authors.forename: F C
authors.initials: FC
authors.lastname: Placido
authors.forename: D N
authors.initials: DN
authors.lastname: Sauder
authors.forename: R A
authors.initials: RA
authors.lastname: Dempsey
authors.forename: P
authors.initials: P
authors.lastname: Amerio
mesh_terms: D002478:Cells, Cultured; D006801:Humans; D007375:Interleukin-1 / Q000096:biosynthesis / Q000235:genetics*; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D008070:Lipopolysaccharides / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology; D012333:RNA, Messenger / Q000032:analysis*; D017472:Receptors, Interleukin-1 / Q000037:antagonists & inhibitors*; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: D007375:Interleukin-1; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D017472:Receptors, Interleukin-1; D011994:Recombinant Proteins