Glycosylation variants of murine interleukin-4: evidence for different functional properties

Immunology. 1992 Jan;75(1):143-9.

Abstract

Immunoprecipitation of biosynthetically labelled interleukin-4 (IL-4) secreted by activated D10 cells yielded three bands after separation under non-reducing conditions by electrophoresis through gradient SDS polyacrylamide gels. This triplet consists of a closely spaced doublet at 23 and 22 kDa (23/22 kDa), and a third band at 18.5 kDa (19 kDa). The 23/22 kDa doublet was converted to the 19 kDa form by enzymatic removal of the N-linked sugars, indicating that the two glycoforms were derived from the 19-kDa core polypeptide. Under reducing conditions, the 19-kDa polypeptide migrated at 14.5 kDa, consistent with the size predicted from the complementary DNA (cDNA). Under non-reducing conditions, IL-4 retained biological activity after electrophoresis and transfer to nitrocellulose. Applying a biological assay, proliferation of the NK cell line, to fractionated nitrocellulose replicas, we found that IL-4 activity was detected over a relatively broad range of molecular weights, reflecting the multiple bands found by immunoprecipitation. This was true not only for IL-4 produced by D10 cells but also for splenic cells activated in vitro. All of the immunoprecipitated IL-4 species were active in inducing proliferation of the NK cell line. However, when the D10 cell line was used to detect IL-4, the 19-kDa species was significantly more active than the higher molecular weight species. These results suggest that different forms of IL-4 may have different functional properties.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Biological Assay
  • Cell Division / immunology
  • Cell Line
  • Cells, Cultured
  • Electrophoresis, Polyacrylamide Gel
  • Glycosylation
  • Immunoblotting
  • Interleukin-4 / chemistry*
  • Interleukin-4 / physiology*
  • Mice
  • Precipitin Tests

Substances

  • Interleukin-4