title: Development of glycosylated human interleukin-1 alpha, neoglyco IL-1 alpha, coupled with D-mannose dimer: synthesis and biological activities in vitro.
abstract: Interleukin 1 (IL-1) is a nonglycosylated cytokine with pleiotropic effects on various cell types. In order to investigate the effect of carbohydrate introduction on IL-1 activity and to develop IL-1 with less deleterious effects recombinant human IL-1 alpha was chemically coupled with mannose dimers, alpha-D-Man1-6-D-Man[Man2(alpha 1,6)] and alpha-D-Man1-4-D-Man[Man2(alpha 1,4)]. About 5 molecules of mannose dimers were introduced per molecule of IL-1. Anti-IL-1 alpha antibody reacted only weakly with the glycosylated IL-1s. Conversely, antibody against the mannose dimer reacted with only glycosylated IL-1. The effect of glycosylation on IL-1 activity was evaluated by measuring a variety of IL-1 activities in vitro, including proliferative effect on T cells, antiproliferative effect on melanoma cells, stimulatory effect on IL-6 synthesis by melanoma cells, and stimulatory effect on prostaglandin E2 synthesis by fibroblast cells. Glycosylated IL-1s exhibited reduced activities, which were 10-fold to more than several hundred-fold lower than those of the original IL-1 alpha depending upon different aspects of activities addressed. Man2(alpha 1,6)-introduced IL-1 exhibited lower activity than Man2(alpha 1,4)-introduced IL-1. The competitive binding of 125I-IL-1 alpha to mouse T cells with unlabeled IL-1s suggests that the reduced activity of glycosylated IL-1s is due, at least partially, to the decrease of their receptor binding abilities.
authors.affiliation: Department of Hygienic Chemistry, Faculty of Pharmaceutical Sciences, Nagoya City University, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Takei
authors.forename: K
authors.initials: K
authors.lastname: Wada
authors.forename: T
authors.initials: T
authors.lastname: Chiba
authors.forename: H
authors.initials: H
authors.lastname: Hayashi
authors.forename: H
authors.initials: H
authors.lastname: Ishihara
authors.forename: K
authors.initials: K
authors.lastname: Onozaki
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002240:Carbohydrate Sequence; D002455:Cell Division / Q000187:drug effects; D002999:Clone Cells; D015232:Dinoprostone / Q000096:biosynthesis; D004187:Disaccharides; D004305:Dose-Response Relationship, Drug; D006023:Glycoproteins / Q000138:chemical synthesis* / Q000494:pharmacology; D006031:Glycosylation; D006801:Humans; D007375:Interleukin-1 / Q000031:analogs & derivatives* / Q000138:chemical synthesis* / Q000494:pharmacology; D008213:Lymphocyte Activation / Q000187:drug effects; D008358:Mannose; D051379:Mice; D008969:Molecular Sequence Data; D009844:Oligosaccharides; D011994:Recombinant Proteins / Q000494:pharmacology; D013329:Structure-Activity Relationship; D013601:T-Lymphocytes; D014407:Tumor Cells, Cultured
chemical_list: D004187:Disaccharides; D006023:Glycoproteins; D007375:Interleukin-1; D009844:Oligosaccharides; D011994:Recombinant Proteins; D015232:Dinoprostone; D008358:Mannose