Journal
JOURNAL OF IMMUNOLOGICAL METHODS
Volume 486, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.jim.2020.112835
Keywords
Cytokines; Inflammation; ELISA; Sheep
Categories
Funding
- National Health and Medical Research Council [APP114576]
- Centre for Research Excellence for Advanced Cardiorespiratory Therapies Improving Organ Support [1079421 - CRE ACTIONS]
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There is growing evidence that inflammation underpins many common diseases. Inflammatory/immunomodulatory/immune mediators, such as cytokines, are key modulators of inflammation and mediate both immune cell recruitment and complex intracellular signalling pathways. Ovine models of disease are increasingly utilized in pre-clinical research, however existing methods for measuring cytokine levels are limited. We established and validated enzyme-linked immunosorbent assays (ELISAs) targeting interleukin (IL)-1 beta, IL-6, IL-8 and IL-10 in sheep plasma. These ELISAs showed high sensitivity and specificity with intra- and inter-assay CVs below 10%, and recovery rates between 82 and 123%. Sensitivity for IL-1 beta, IL-6, IL-8 and IL-10 were 117.6 pg/mL, 443.1 pg/mL, 30.9 pg/mL, and 64.3 pg/mL, respectively. ELISA test result reproducibility decreased significantly after 12 weeks of plasma storage at -80 degrees C. Therefore, for accurate cytokine measurements, plasma samples need to be tested within three months of sample collection to account for cytokine protein degradation. These ELISAs offer a reliable and convenient method to identify inflammatory cytokine changes in sheep, allowing key insights into the disease pathogenesis of these ruminants.
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