Detection of anti-TNFα activity in canine hyperimmune serum using a TNFα inhibition assay

Kotiw, Michael, Morgan, Michael, Taylor, Stephen M. and Shiels, Ian A. (2009) Detection of anti-TNFα activity in canine hyperimmune serum using a TNFα inhibition assay. Veterinary Clinical Pathology, 39 1: 46-52. doi:10.1111/j.1939-165X.2009.00166.x

Author Kotiw, Michael
Morgan, Michael
Taylor, Stephen M.
Shiels, Ian A.
Title Detection of anti-TNFα activity in canine hyperimmune serum using a TNFα inhibition assay
Formatted title
Detection of anti-TNFα activity in canine hyperimmune serum using a TNFα inhibition assay
Journal name Veterinary Clinical Pathology   Check publisher's open access policy
ISSN 0275-6382
Publication date 2009-07
Year available 2009
Sub-type Article (original research)
DOI 10.1111/j.1939-165X.2009.00166.x
Volume 39
Issue 1
Start page 46
End page 52
Total pages 7
Editor Karen M. Young
Place of publication United States
Publisher Wiley-Blackwell
Collection year 2010
Language eng
Subject 0707 Veterinary Sciences
070705 Veterinary Immunology
Formatted 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.
Keyword Dog
Escherichia coli J5
hyperimmune serum
L929 cell assay
TNFα inhibition
Q-Index Code C1
Q-Index Status Confirmed Code
Additional Notes Early View (Articles online in advance of print)- Published Online: 1 Jul 2009

Document type: Journal Article
Sub-type: Article (original research)
Collections: 2010 Higher Education Research Data Collection
School of Veterinary Science Publications
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Citation counts: TR Web of Science Citation Count  Cited 1 times in Thomson Reuters Web of Science Article | Citations
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Created: Mon, 16 Nov 2009, 14:43:49 EST by Maria Campbell on behalf of School of Veterinary Science