Specific human granulocyte-macrophage colony-stimulating factor antagonists

Timothy R. Hercus, Christopher J. Bagley, Bronwyn Cambareri, Mara Dottore, Joanna M. Woodcock, Mathew A. Vadas, M. Frances Shannon, Angel F. Lopez

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101 Citations (Scopus)

Abstract

Human granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic hemopoietic growth factor and activator of mature myeloid cell function. We have previously shown that residue 21 in the first helix of GM- CSF plays a critical role in both biological activity and high-affinity receptor binding. We have now generated analogues of GM-CSF mutated at residue 21, expressed them in Escherichia coli, and examined them for binding, agonistic, and antagonistic activities. Binding experiments showed that GM E21A, E21Q, E21F, E21H, E21R, and E21K bound to the GM-CSF receptor α chain with a similar affinity to wild-type GM-CSF and had lost high- affinity binding to the GM-CSF receptor α-chain-common β-chain complex. From these mutants, only the charge reversal mutants E21R and E21K were completely devoid of agonistic activity. Significantly we found that E21R and E21K antagonized the proliferative effect of GM-CSF on the erythroleukemic cell line TF-1 and primary acute myeloid leukemias, as well as GM-CSF- mediated stimulation of neutrophil superoxide production. This antagonism was specific for GM-CSF in that no antagonism of interleukin 3-mediated TF-1 cell proliferation or tumor necrosis factor α-mediated stimulation of neutrophil superoxide production was observed. E. coli-derived GM E21R and E21K were effective antagonists of both nonglycosylated and glycosylated wild-type GM- CSF. These results show that low-affinity GM-CSF binding can be dissociated from receptor activation and have potential clinical significance for the management of inflammatory diseases and certain leukemias where GM-CSF plays a pathogenic role.

Original languageEnglish
Pages (from-to)5838-5842
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume91
Issue number13
DOIs
Publication statusPublished or Issued - 21 Jun 1994

Keywords

  • cytokines
  • inflammation
  • leukemia
  • receptors

ASJC Scopus subject areas

  • General

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