Distinct reactivities of interleukin-4-specific antibodies with recombinant and native canine interleukin-4 in various assays.
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.
Lauber
Beatrice
B
Department of Clinical Research, Vetsuisse Faculty, University of Bern, Switzerland.
Marti
Eliane
E
Plattet
Philippe
P
Urwyler
Adrian
A
Rai
Muhammad Farooq
MF
Schmidt
Michael F G
MF
Welle
Monika
M
Zurbriggen
Andreas
A
Janda
Jozef
J
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D015153:Blotting, Western; D002460:Cell Line; D004285:Dogs / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D015847:Interleukin-4 / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology
D000906:Antibodies; D011994:Recombinant Proteins; D015847:Interleukin-4