Journal article
Development of detection methods for ruminant interleukin (IL)-4
Journal of immunological methods, Vol.301(1), pp.114-123
2005
Handle:
https://hdl.handle.net/2376/109078
PMID: 15979636
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.
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Details
- Title
- Development of detection methods for ruminant interleukin (IL)-4
- Creators
- J.C Hope - Institute for Animal Health, Compton, Near Newbury, RG20 7NN, UKL.S Kwong - Institute for Animal Health, Compton, Near Newbury, RG20 7NN, UKM Thom - Institute for Animal Health, Compton, Near Newbury, RG20 7NN, UKP Sopp - Institute for Animal Health, Compton, Near Newbury, RG20 7NN, UKW Mwangi - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USAW.C Brown - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USAG.H Palmer - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USAS Wattegedera - Moredun Research Institute, International Research Centre, Pentlands Science Park, Bush Loan, Edinburgh, EH26 0PZ, UKG Entrican - Moredun Research Institute, International Research Centre, Pentlands Science Park, Bush Loan, Edinburgh, EH26 0PZ, UKC.J Howard - Institute for Animal Health, Compton, Near Newbury, RG20 7NN, UK
- Publication Details
- Journal of immunological methods, Vol.301(1), pp.114-123
- Academic Unit
- Veterinary Microbiology and Pathology, Department of; Paul G. Allen School for Global Animal Health
- Publisher
- Elsevier B.V
- Identifiers
- 99900547169201842
- Language
- English
- Resource Type
- Journal article