Development of glycosylated human interleukin-1alpha, neoglyco IL-1alpha, by coupling with D-galactose monosaccharide: biological activities in vitro.
In the previous study, galactose with C9 spacer was chemically coupled to human recombinant (rh) IL-1alpha in order to study the effect of glycosylation on its activities, and to develop IL-1 with less deleterious effects. In this study we examined a variety of IL-1 activities in vitro, including proliferative effect on T cells, antiproliferative effect on myeloid leukemic cells and melanoma cells, stimulatory effects on IL-6 synthesis by melanoma cells and PGE2 synthesis by fibroblast cells Galactose-introduced IL-1alpha (Gal-IL-1alpha) exhibited reduced activities from 10 to 10000 times compared with unmodified IL-1alpha in all the activities performed in vitro. The competitive binding of 125I-IL-1alpha to mouse T cells and pre-B cells with unlabeled IL-1alpha s suggests a decrease in binding affinities of Gal-IL-1alpha to both type I and type II IL-1 receptors. Therefore, reduced activities of Gal-IL-1alpha are due, at least partially, to the decrease in their receptor binding affinities.
Nabeshima
S
S
Department of Hygienic Chemistry, Faculty of Pharmaceutical Sciences, Nagoya City University, Tanabe, Mizuho, Japan. konozaki@phar.nagoya-cu.ac.jp
Chiba
T
T
Takei
Y
Y
Watanabe
S
S
Okuyama
H
H
Onozaki
K
K
D000818:Animals; D001667:Binding, Competitive; D015232:Dinoprostone / Q000096:biosynthesis; D005690:Galactose; D006023:Glycoproteins / Q000502:physiology; D006801:Humans; D007375:Interleukin-1 / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D015850:Interleukin-6 / Q000096:biosynthesis; D007951:Leukemia, Myeloid / Q000473:pathology; D008213:Lymphocyte Activation; D008545:Melanoma / Q000473:pathology; D051379:Mice; D011994:Recombinant Proteins; D013329:Structure-Activity Relationship; D013601:T-Lymphocytes / Q000187:drug effects; D014407:Tumor Cells, Cultured
D006023:Glycoproteins; D007375:Interleukin-1; D015850:Interleukin-6; D011994:Recombinant Proteins; D015232:Dinoprostone; D005690:Galactose