You are here
Development of detection methods for ruminant interleukin (IL)-4.
Title | Development of detection methods for ruminant interleukin (IL)-4. |
Publication Type | Journal Article |
Year of Publication | 2005 |
Authors | Hope, JC, Kwong, LS, Thom, M, Sopp, P, Mwangi, W, Brown, WC, Palmer, GH, Wattegedera, S, Entrican, G, Howard, CJ |
Journal | J Immunol Methods |
Volume | 301 |
Issue | 1-2 |
Pagination | 114-23 |
Date Published | 2005 Jun |
ISSN | 0022-1759 |
Keywords | Animals, Antibodies, Monoclonal, Antigens, Cattle, Cercopithecus aethiops, COS Cells, Enzyme-Linked Immunosorbent Assay, Interleukin-4, Th1 Cells, Th2 Cells |
Abstract | 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. |
DOI | 10.1016/j.jim.2005.04.010 |
Alternate Journal | J. Immunol. Methods |
PubMed ID | 15979636 |