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Development and application of specific cytokine assays in tissue samples from a bottlenose dolphin with hyperinsulinemia
- Eberle, Kirsten C., Waters, Theresa E., Jensen, Eric D., Venn-Watson, Stephanie K., Sacco, Randy E.
- Frontiers in endocrinology 2013 v.4 pp. 134
- Tursiops truncatus, antibodies, biomarkers, case studies, dolphins, fatty liver, fluorescence, gene expression, gene expression regulation, humans, hyperglycemia, hyperinsulinemia, immunohistochemistry, inflammation, insulin resistance, interferon-gamma, interleukin-10, interleukin-12, interleukin-13, interleukin-18, interleukin-1beta, interleukin-2, interleukin-4, interleukin-6, interleukin-8, leukocyte count, liver, males, messenger RNA, noninsulin-dependent diabetes mellitus, quantitative polymerase chain reaction, spleen, transforming growth factor beta, tumor necrosis factor-alpha
- Chronic inflammation has been associated with insulin resistance and type 2 diabetes (T2D) in humans. Postmortem hepatic and splenic tissue from a 46-year old geriatric male bottlenose dolphin (Tursiops truncatus) with insulin resistance (chronic hyperinsulinemia with hyperglycemia), chronic inflammation (white blood cell count greater than 12,000 cells/µl), and mild fatty liver disease was evaluated for elevated pro-inflammatory mediators. Cytokine mRNA expression in postmortem hepatic and splenic tissue, as determined by real-time PCR, included an array of cytokines: TGF-ß, TNF-α, IFN-γ, IL-2, IL-4, IL-10, IL-12p40, IL-13, and IL-18. Values from this dolphin were compared to a younger reference dolphin with no known chronic metabolic perturbations or inflammation. Levels of TGF-ß,TNF-α, and IL-4 were higher in the case dolphin's liver compared to that of the reference dolphin. In the case dolphin's spleen, IL-10 and IFN-γ mRNA was upregulated while IL-4 was less than reference dolphin.. IL-18 and IL-13 were upregulated in both tissues. Fluorescent immunohistochemistry (IHC) utilized the following antibodies: anti-porcine IL-6, anti-bovine IFN-γ, IL-4, and IL-10, anti-human TGF-β, anti-ovine IL-1β, and anti-dolphin IL-8. Fluorescent IHC in spleen from the case dolphin revealed staining of IL-4, IL-6, IL-8 and TGF-β throughout the tissue. IL-10 and IFN-γ were seen to predominatein areas surrounding the follicles of splenic tissue. This is the first characterization of cytokine levels in dolphin hepatic and splenic tissue. While there are limitations to a case study, this report of inflammatory biomarkers in tissues of a dolphin with insulin resistance and fatty liver disease are similar to those observed in human patients.