Detection of anti-TNFα activity in canine hyperimmune serum using a TNFα inhibition assay
Article
Kotiw, Michael, Morgan, Michael, Taylor, Stephen M. and Shiels, Ian A.. 2010. "Detection of anti-TNFα activity in canine hyperimmune serum using a TNFα inhibition assay
." Veterinary Clinical Pathology. 39 (1), pp. 46-52. https://doi.org/10.1111/j.1939-165X.2009.00166.x
| Article Title | Detection of anti-TNFα activity in canine hyperimmune serum using a TNFα inhibition assay |
|---|---|
| ERA Journal ID | 5553 |
| Article Category | Article |
| Authors | Kotiw, Michael (Author), Morgan, Michael (Author), Taylor, Stephen M. (Author) and Shiels, Ian A. (Author) |
| Journal Title | Veterinary Clinical Pathology |
| Journal Citation | 39 (1), pp. 46-52 |
| Number of Pages | 7 |
| Year | 2010 |
| Place of Publication | Hoboken, NJ. United States |
| ISSN | 0275-6382 |
| 1939-165X | |
| Digital Object Identifier (DOI) | https://doi.org/10.1111/j.1939-165X.2009.00166.x |
| Abstract | Background: Increased serum tumor necrosis factor-α (TNFα) activity has been associated with onset of serious inflammatory diseases in dogs. Development of treatment with TNFα-antagonists has been limited by the unavailability of suitable reagents and potency assays for TNFα. Objectives: The objectives of this study were to optimize a cell-based assay to measure anti-TNFα activity in serum and plasma from hyperimmune (vaccinated with an Escherichia coli J5 bacterin) and unvaccinated canine donors; to use the assay to determine whether hyperimmune serum inhibits TNFα activity in vivo; and to determine whether soluble TNF receptor-1 (sTNFR1, a naturally occurring TNFα antagonist) contributes to anti-TNFα activity. Methods: Commercial plasma and serum from hyperimmune-frozen plasma (HFP) donors and unvaccinated fresh-frozen plasma (FFP) donors were used in the study. An L929-cell TNFα-inhibition assay (LTIA) was optimized to measure anti-TNFα activity. Using a rat subcutaneous pouch model of inflammation, the effects of HFP, FFP, a synthetic TNFα antagonist (Etanercept), and carprofen on TNFα activity were compared in vivo. Immunofluorescence was used to measure soluble sTNFR1 concentration. Results: Using the optimized LTIA, HFP serum but not FFP serum decreased canine TNFα activity (P<.01). HFP plasma and Etanercept (but not FFP plasma or carprofen) significantly decreased TNFα activity in pouch exudates (P<.05). A significantly higher concentration of sTNFR1 was found in HFP than FFP serum. Conclusions: Using the LTIA, anti-TNFα activity is readily measured in canine serum and inflammatory exudates. sTNFR1 appears to contribute to anti-TNFα activity in HFP serum. These results suggest HFP should be investigated further as a potential immunotherapeutic agent for controlling canine diseases in which TNFα is implicated. |
| Keywords | dogs; escherichia coli J5; hyperimmune serum; L929 cell assay; TNF inhibition |
| ANZSRC Field of Research 2020 | 320402. Applied immunology (incl. antibody engineering, xenotransplantation and t-cell therapies) |
| 310701. Bacteriology | |
| 300906. Veterinary immunology | |
| Public Notes | Files associated with this item cannot be displayed due to copyright restrictions. |
| Byline Affiliations | Department of Biological and Physical Sciences |
| University of Queensland |
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