Specific inhibition of interleukin 3 bioactivity by a monoclonal antibody reactive with hematopoietic progenitor cells.
HIM1, originally designated HI98, a murine monoclonal IgM antibody raised against human mononuclear cells, has been reported at the Fourth International Leukocyte Typing Workshop (called antibody M0141) to be the only one of 157 antibodies tested that inhibited binding of interleukin 3 (IL-3) to KG-1 human acute myelogenous leukemia cells and normal human monocytes. We have carried out detailed studies of the selective effect of HIM1 on IL-3-mediated stimulation of hematopoietic progenitors. Preincubation of normal human bone marrow mononuclear cells, depleted of adherent cells and T cells, with HIM1 antibody resulted in a dose-dependent inhibition of IL-3-mediated stimulation of both erythroid burst-forming units (maximum inhibition 55%) and granulocyte/macrophage colony-forming units (maximum inhibition 49%). HIM1 antibody had no effect on growth of erythroid colony-forming units in culture. In addition, preincubation of the cells with HIM1 antibody had no deleterious effect on granulocyte/macrophage colony-stimulating factor-induced growth of either erythroid bursts or granulocyte/macrophage colonies. To be certain that the HIM1 antibody did not react directly with IL-3 itself, we attempted to use immunodepletion to remove IL-3 that had been added to our culture medium. Although we were able to remove IL-3 bioactivity by immunodepletion with anti-IL-3 antibody bound to Sepharose beads, beads with attached HIM1 did not remove IL-3 activity from the medium. Polymorphonuclear neutrophils bind high levels of HIM1, although they have very few or no detectable IL-3 receptors. Therefore, this antibody appears to recognize a cell surface antigen that is critical for optimal IL-3 binding and bioactivity but is not the actual IL-3 receptor.
Emanuel
P D
PD
Department of Medicine, University of Alabama, Birmingham 35294.
Peiper
S C
SC
Chen
Z
Z
Sheng
D C
DC
Zuckerman
K S
KS
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D001665:Binding Sites; D002478:Cells, Cultured; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000276:immunology*; D006801:Humans; D007377:Interleukin-3 / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D007700:Kinetics
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D007377:Interleukin-3