Production of antibodies to canine IL-1beta and canine TNF to assess the role of proinflammatory cytokines

Baggio, Vannozza; Ott, Fabienne; Fischer, René W.; Gram, Hermann; Peele, John; Spreng, David; Schmökel, Hugo; Jungi, Thomas (2005). Production of antibodies to canine IL-1beta and canine TNF to assess the role of proinflammatory cytokines. Veterinary immunology and immunopathology, 107(1-2), pp. 27-39. Elsevier 10.1016/j.vetimm.2005.02.019

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IL-1 and TNF are important proinflammatory cytokines implicated in both antimicrobial host defense and pathogenesis of diseases with an immune-mediated and/or inflammatory component. Respective studies in the dog have been hampered by the unavailability of reagents allowing the specific measurement of canine cytokine proteins and the effect of canine cytokine neutralization by Ab. Starting with recombinant canine (rcan) IL-1beta and rcanTNF, four polyclonal antisera and 22 mAb specific for rcanIL-1beta and rcanTNF were generated. Their usefulness in neutralization assays was determined. Using cytokine-containing supernatants of canine cells in bioassays, polyclonal antisera neutralized either canine IL-1beta or TNF. TNF was also neutralized by three antibodies developed in this study and one commercial mAb. The usefulness of monoclonal and polyclonal Ab in canine cytokine-specific Ab capture ELISA's was assessed. This resulted in the identification of a commercial mAb combination and one pair developed in this study allowing low levels of TNF to be detected by antibody capture ELISA. The detection limit was 141 pg/ml rcanTNF for both combinations. Using rcanIL-1beta as an antigen allowed the detection of lower concentrations of rcanIL-1beta (20 pg/ml, on the average) by a pair of polyclonal antisera than when monoclonals were used. By using such IL-1beta-specific and TNF-specific ELISA's, the respective cytokines were detected in supernatants of canine PBMC stimulated with LPS or heat-killed Listeria monocytogenes and interferon-gamma combined. Thus, monoclonal and polyclonal reagents were identified allowing the quantitation of canine IL-1beta and TNF production in vitro, and the neutralization of these cytokines.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Department of Clinical Veterinary Medicine (DKV) > Small Animal Clinic > Small Animal Clinic, Surgery
05 Veterinary Medicine > Department of Clinical Veterinary Medicine (DKV) > Small Animal Clinic
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Virology and Immunology

UniBE Contributor:

Spreng, David; Schmökel, Hugo and Jungi, Thomas

Subjects:

600 Technology > 630 Agriculture

ISSN:

0165-2427

Publisher:

Elsevier

Language:

English

Submitter:

Simone Forterre

Date Deposited:

24 Sep 2014 15:09

Last Modified:

25 Dec 2014 01:12

Publisher DOI:

10.1016/j.vetimm.2005.02.019

PubMed ID:

15982477

Uncontrolled Keywords:

IL-1b, TNF, Ab to canine cytokines, Ab to cytokines, Ab, Polyclonal, mAb, Dog

BORIS DOI:

10.7892/boris.58577

URI:

https://boris.unibe.ch/id/eprint/58577

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