Distinct reactivities of interleukin-4-specific antibodies with recombinant and native canine interleukin-4 in various assays

Beatrice Lauber, Eliane Marti, Philippe Plattet, Adrian Urwyler, Muhammad Farooq Rai, Michael F.G. Schmidt, Monika Welle, Andreas Zurbriggen, Jozef Janda

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Interleukin 4 (IL-4) plays a central role in immune responses to parasites and allergens. IL-4 drives the differentiation of naive T cells into Th2 cells and regulates immunoglobulin class switching to IgE.Little is known about the role of IL-4 in canine allergies and parasite infections. Most of the information derives from measurement of IL-4 mRNA expression in dog tissues, but detection of IL-4 protein has been difficult so far, probably due to low sensitivity of available methods. Antibodies (Ab) specific for canine IL-4 are available from various sources, but these Ab have been produced against recombinant Escherichia coli-expressed canine IL-4 and there is only limited information on their reactivities with native canine IL-4. Therefore, in the present study, we tested six available canine IL-4-specific Ab for their reactivities with recombinant canine IL-4 expressed in E. coli (rec.IL-4) or in mammalian cells (mam.IL-4), and with supernatants from stimulated canine peripheral blood mononuclear cells (PBMCs) using several detection methods, including Western blotting, ELISA, cytokine bead assay, and intracellular IL-4 staining. Additionally, we tested a bovine IL-4-specific antibody that has been previously shown to cross-react with canine IL-4. All tested Ab except anti-bovine IL-4 reacted with rec.IL-4, and most of them reacted with mam.IL-4. However, only the cytokine bead assay was sensitive enough to allow the detection of IL-4 in supernatants of canine PBMCs.

Original languageEnglish
Pages (from-to)310-316
Number of pages7
JournalVeterinary Immunology and Immunopathology
Volume137
Issue number3-4
DOIs
StatePublished - Oct 2010

Keywords

  • Antibody
  • Canine
  • Cytokine bead assay
  • Interleukin-4

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