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. Determinations of plasma cytokine concentrations —Concentrations of proinflammatory (IL-1β, TNF-α, IL-6, and IL-17A) and anti-inflammatory (IL-10 and IL-13) cytokines were measured in plasma samples with specific ELISA commercial kits a according to the

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in American Journal of Veterinary Research

diagnostic tests, it is worthwhile to evaluate the use of inflammatory mediators as potential biomarkers. In humans with asthma, increases in concentrations of IL-4, TNF-α, and NO metabolites or exhaled NO have been found, prompting interest into further

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in American Journal of Veterinary Research

equine asthma have an increase in the expression of the proinflammatory cytokine TNF-α and the chemokine IL-8 and a decrease in expression of an anti-inflammatory cytokine IL-10, compared with expression in horses without respiratory disease

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in American Journal of Veterinary Research

in basal cell culture media (Dulbecco modified Eagle medium–Ham F12, 1:1) devoid of hormonal and growth factor supplements. The following recombinant equine cytokines were used: TNF-α, IL-2, IL-4, IL-5, IL-6, IL-8, and IL-10. k,l After 48 hours

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in American Journal of Veterinary Research

−80°C until extraction of RNA. Cytokine analysis —Cytokine mRNA (IL-1A, IL1B, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, interferon-G, and TNF-α) was quantitated by use of a real-time quantitative reverse-transcription PCR assay and species

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in American Journal of Veterinary Research

set for equine TNF-α was designed according to Giguère et al. 31 Reference genes were GAPDH, RPL-32, HPRT, and SDHA. Target genes were innate inflammatory (TNF-α and IL-8), Th1 cell–derived (interferon-γ and IL-1β), Th2 cell–derived (IL-4, IL-5, IL-6

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in American Journal of Veterinary Research

proinflammatory cytokines and chemokines, including IL-8, G-CSF, and CXCL1. 14,16 In addition, IL-17 induces the synthesis of TNF-α and IL-1β from macrophages, and these 2 cytokines, along with interferon-γ, enhance the effects of IL-17 on airway epithelium. 12

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in American Journal of Veterinary Research