Monoclonal antibodies against the human interleukin-11 receptor alpha-chain (IL-11Ralpha) and their use in studies of human mononuclear cells

J Immunol Methods. 2000 Jul 31;241(1-2):43-59. doi: 10.1016/s0022-1759(00)00194-0.

Abstract

A panel of 14 hybridoma cell lines secreting monoclonal antibodies against the human interleukin-11 receptor alpha chain (hIL-11Ralpha) was obtained using two different approaches. Two antibodies were raised against peptides of the N- and C-terminal sequences, respectively, of the extracellular part of the hIL-11Ralpha. Another group of 12 antibodies was generated against a hybrid protein consisting of the extracellular part of the hIL-11Ralpha fused to mature full-length human IL-2. All these antibodies recognized native hIL-11Ralpha and most also recognized the denatured receptor on immunoblots after SDS-PAGE. Four different epitopes were identified on the extracellular part of the hIL-11Ralpha. One epitope, defined by the E27 antibody, is located at the N-terminus and the other three epitopes are clustered in the membrane-proximal, C-terminal region. The antibodies defining epitopes I and II recognized membrane-bound hIL-11Ralpha expressed in gp130/hIL-11Ralpha-co-transfected Ba/F3 cells. The E27 antibody cross-reacted with murine IL-11Ralpha, in agreement with the fact that the N-terminal region is highly conserved between species. The other 13 antibodies all recognized a region between amino acids 319 and 363, which is the membrane-proximal part of the hIL-11Ralpha. This region, which is less conserved between mouse and human, is shown here to be an immunodominant region. Anti-IL-11Ralpha monoclonal antibodies, which have not been described previously enabled us to explore the expression and tissue distribution of IL-11Ralpha on human peripheral blood mononuclear cells and cell lines. The antibodies provide powerful tools for the study of the regulation and function of the receptor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal*
  • Antibody Affinity
  • Cross Reactions
  • Epitopes
  • Flow Cytometry / methods*
  • Humans
  • Interleukin-11 Receptor alpha Subunit
  • Interleukin-2 / genetics
  • Interleukin-2 / immunology
  • Leukocytes, Mononuclear*
  • Mice
  • Molecular Sequence Data
  • Peptide Fragments / immunology
  • Receptors, Interleukin / genetics
  • Receptors, Interleukin / immunology*
  • Receptors, Interleukin / isolation & purification*
  • Receptors, Interleukin-11
  • Recombinant Fusion Proteins / immunology
  • Sequence Homology, Amino Acid
  • Species Specificity
  • Surface Plasmon Resonance
  • Tissue Distribution

Substances

  • Antibodies, Monoclonal
  • Epitopes
  • IL11RA protein, human
  • Il11ra1 protein, mouse
  • Il11ra2 protein, mouse
  • Interleukin-11 Receptor alpha Subunit
  • Interleukin-2
  • Peptide Fragments
  • Receptors, Interleukin
  • Receptors, Interleukin-11
  • Recombinant Fusion Proteins