Development of detection methods for ruminant interleukin (IL)-4.

Recombinant bovine IL-4 (rbo IL-4) was transiently expressed in COS-7 cells. Mice were immunised with a plasmid encoding rbo IL-4 and boosted with rbo IL-4. A number of monoclonal antibodies (mAb) were generated that reacted with rbo IL-4 in an ELISA and these cloned hybridomas were termed CC311, CC312, CC313 and CC314. A pair of mAb (CC313 and CC314) was identified that together could be used to detect both recombinant and native bovine IL-4 by ELISA and a luminometric detection method was applied to the ELISA. Using this method native bovine IL-4 was detected in supernatants of PBMC stimulated with mitogens. In addition, high level secretion of IL-4 by Fasciola hepatica specific Th2 clones, but not by a Babesia bovis specific Th1 clone, was confirmed. The ELISA was also able to detect recombinant ovine IL-4. The pair of mAb used for ELISA could also be used for the detection of IL-4 spot forming cells by ELISPOT. In addition intracytoplasmic expression of IL-4 could be detected. The ability to detect ruminant IL-4 by three methods: ELISA, ELISPOT and by flow cytometric analysis of intracytoplasmic expression will permit studies of the role of this important cytokine in the immunology and pathogenesis of animal diseases.
AuthorsJ C Hope, L S Kwong, M Thom, P Sopp, W Mwangi, W C Brown, G H Palmer, S Wattegedera, G Entrican, C J Howard
JournalJournal of immunological methods (J Immunol Methods) Vol. 301 Issue 1-2 Pg. 114-23 (Jun 2005) ISSN: 0022-1759 [Print] Netherlands
PMID15979636 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Antibodies, Monoclonal
  • Antigens
  • Interleukin-4
  • Animals
  • Antibodies, Monoclonal (immunology)
  • Antigens (immunology)
  • COS Cells
  • Cattle (immunology)
  • Cercopithecus aethiops
  • Enzyme-Linked Immunosorbent Assay
  • Interleukin-4 (analysis, immunology)
  • Th1 Cells (secretion)
  • Th2 Cells (secretion)

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