Identification of a distinct low-affinity receptor for human interleukin-4 on pre-B cells

Blood. 1993 Jun 1;81(11):2998-3005.

Abstract

Biotinylated interleukin-4 (IL-4) was used to examine IL-4 receptor (IL-4R) expression on a range of human B-cell lines by flow cytometry. Using high concentrations of biotinylated IL-4, we have identified a novel low-affinity IL-4 receptor expressed at high levels on pre-B lines. Expression of this low-affinity receptor did not correlate with detected mRNA levels for the previously cloned receptor or with reactivity of two anti-human IL-4R monoclonal antibodies (MoAb). Radiolabeled IL-4 cross-linking studies using pre-B lines showed a doublet of 65 to 75 Kd in contrast to the 110- to 130-Kd molecule detected on cells expressing the cloned IL-4R. A soluble IL-4 binding protein (IL-4bp) was purified from the supernatants of three pre-B lines expressing the low-affinity receptor on their surface. IL-4bp could block both IL-4-mediated CD23 induction on tonsil B cells and IL-4-induced inhibition of proliferation of the pre-B line JM1. Partial N-terminal amino acid sequence was obtained from purified IL-4bp that confirmed this protein to be novel. A 12 amino acid peptide based on the IL-4bp sequence was used to produce a polyclonal antiserum that was reactive with purified IL-4bp, and also bound to the surface of pre-B cells but not to murine CTLL cells transfected with the human IL-4R. Blocking MoAb against the previously characterized high-affinity receptor inhibited IL-4-mediated proliferation of hIL-4R+ CTLL cells but had no effect on IL-4-induced inhibition of JM1 cell proliferation, and only partially inhibited IL-4-mediated CD23 and sIgM induction and proliferation of tonsil B cells. The data presented here provide evidence for a novel cell-surface expressed low-affinity IL-4R that also exists as a biologically active soluble IL-4 binding protein.

MeSH terms

  • Amino Acid Sequence
  • Antibodies, Monoclonal / immunology
  • B-Lymphocytes / chemistry*
  • Cross-Linking Reagents
  • Gene Expression
  • Humans
  • Interleukin-4 / metabolism*
  • Molecular Sequence Data
  • RNA, Messenger / genetics
  • Receptors, Interleukin-4
  • Receptors, Mitogen / chemistry*
  • Receptors, Mitogen / immunology
  • Solubility

Substances

  • Antibodies, Monoclonal
  • Cross-Linking Reagents
  • RNA, Messenger
  • Receptors, Interleukin-4
  • Receptors, Mitogen
  • Interleukin-4