In vitro and in vivo characterisation of anti-murine IL-13 antibodies recognising distinct functional epitopes

Int Immunopharmacol. 2009 Feb;9(2):201-6. doi: 10.1016/j.intimp.2008.11.001. Epub 2008 Nov 27.

Abstract

Interleukin-13 (IL-13) sequentially binds to IL-13Ralpha1 and IL-4Ralpha forming a high affinity signalling complex. This receptor complex is expressed on multiple cell types in the airway and signals through signal transducer and activator of transcription factor-6 (STAT-6) to stimulate the production of chemokines, cytokines and mucus. Antibodies have been generated, using the UCB Selected Lymphocyte Antibody Method (UCB SLAM), that block either binding of murine IL-13 (mIL-13) to mIL-13Ralpha1 and mIL-13Ralpha2, or block recruitment of mIL-4Ralpha to the mIL-13/mIL-13Ralpha1 complex. Monoclonal antibody (mAb) A was shown to bind to mIL-13 with high affinity (K(D) 11 pM) and prevent binding of mIL-13 to mIL-13Ralpha1. MAb B, that also bound mIL-13 with high affinity (K(D) 8 pM), was shown to prevent recruitment of mIL-4Ralpha to the mIL-13/mIL-13Ralpha1 complex. In vitro, mAbs A and B similarly neutralised mIL-13-stimulated STAT-6 activation and TF-1 cell proliferation. In vivo, mAbs A and B demonstrated equipotent, dose-dependent inhibition of eotaxin generation in mice stimulated by intraperitoneal administration of recombinant mIL-13. In an allergic lung inflammation model in mice, mAbs A and B equipotently inhibited muc5ac mucin mRNA upregulation in lung tissue measured two days after intranasal allergen challenge. These data support the design of therapeutics for the treatment of allergic airway disease that inhibits assembly of the high affinity IL-13 receptor signalling complex, by blocking the binding of IL-13 to IL-13Ralpha1 and IL-13Ralpha2, or the subsequent recruitment of IL-4Ralpha.

MeSH terms

  • Animals
  • Antibodies, Blocking / immunology*
  • Antibodies, Monoclonal / immunology*
  • Cell Line
  • Cell Line, Tumor
  • Chemokine CCL11 / analysis
  • Chemokine CCL11 / immunology
  • Disease Models, Animal
  • Epitopes / immunology
  • Humans
  • Hypersensitivity / immunology
  • Interleukin-13 / antagonists & inhibitors*
  • Interleukin-13 / immunology
  • Interleukin-13 Receptor alpha1 Subunit / antagonists & inhibitors*
  • Interleukin-13 Receptor alpha1 Subunit / immunology
  • Interleukin-13 Receptor alpha2 Subunit / antagonists & inhibitors*
  • Interleukin-13 Receptor alpha2 Subunit / immunology
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mucin 5AC / immunology
  • Mucin 5AC / metabolism
  • Ovalbumin / immunology
  • Pneumonia / immunology
  • Pneumonia / metabolism
  • Rabbits
  • Receptors, Cell Surface / antagonists & inhibitors*
  • Receptors, Cell Surface / immunology
  • Recombinant Proteins / antagonists & inhibitors
  • Recombinant Proteins / immunology
  • Recombinant Proteins / metabolism
  • STAT6 Transcription Factor / immunology
  • STAT6 Transcription Factor / metabolism

Substances

  • Antibodies, Blocking
  • Antibodies, Monoclonal
  • Ccl11 protein, mouse
  • Chemokine CCL11
  • Epitopes
  • Il4ra protein, mouse
  • Interleukin-13
  • Interleukin-13 Receptor alpha1 Subunit
  • Interleukin-13 Receptor alpha2 Subunit
  • Muc5ac protein, mouse
  • Mucin 5AC
  • Receptors, Cell Surface
  • Recombinant Proteins
  • STAT6 Transcription Factor
  • Stat6 protein, mouse
  • Ovalbumin