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.