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

J C Hope; L S Kwong; M Thom; P Sopp; W Mwangi; W C Brown; G H Palmer; S Wattegedera; G Entrican; C J Howard

doi:10.1016/j.jim.2005.04.010

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

J C Hope·L S Kwong·M Thom·P Sopp·W Mwangi·W C Brown·G H Palmer·S Wattegedera·G Entrican·C J Howard

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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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Animals Antibodies, Monoclonal Antigens COS Cells Cattle Chlorocebus aethiops Enzyme-Linked Immunosorbent Assay Interleukin-4 Th1 Cells Th2 Cells Journal Article Research Support, Non-U.S. Gov't

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J C Hope, L S Kwong, M Thom, P Sopp, W Mwangi, W C Brown, G H Palmer, S Wattegedera, G Entrican, C J Howard. 2005. Development of detection methods for ruminant interleukin (IL)-4.. https://doi.org/10.1016/j.jim.2005.04.010

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