Type of Document	Master's Thesis
Author	Sykes, Benjamin William
URN	etd-06172003-143114
Title	Evaluation of Pgg-Glucan, a Novel Immunomodulator, in in Vitro and Ex Vivo Models of Equine Endotoxemia
Degree	Master of Science
Department	Veterinary Medical Sciences
Advisory Committee	Advisor Name Title Furr, Martin O. Committee Chair Donaldson, Lydia L. Committee Member McKenzie, Harold C. III Committee Member
Keywords	horse endotoxemia PGG-Glucan
Date of Defense	2003-06-16
Availability	unrestricted
Abstract Justification - Endotoxemia is an important contributor to mortality and loss of use in the horse and results in significant losses to the equine industry on an annual basis. Objective - To determine the effect of PGG-Glucan on the cytokine response to endotoxin in the horse. Animals - Part 1; 6 adult horses. Part 2; 12 adult horses. Procedure - Part 1; Whole blood was collected, aliquoted, and incubated in vitro in four groups; saline control, endotoxin (LPS) (100 ng/ml), PGG-Glucan (0.1, 1.0, 10 and 100 μg/ml) and LPS (100 ng/ml) plus PGG-Glucan (0.1, 1.0, 10 and 100 μgg/ml). Supernatants were collected at 0, 6 and 12 hours and assayed for tumor necrosis factor £\ (TNF£\) activity. Part 2; Horses received either PGG-Glucan (1 mg/kg) or an equal volume of isotonic saline (0.9% NaCl) IV over 15 minutes. Twenty four hours later blood was collected and mononuclear cells isolated for cell culture. Cells were treated with LPS (100 ng/ml) and RNA extractions were performed at 0, 6, 12, 24 and 48 hours. Relative mRNA expression of TNFα, interleukin-1β (IL-1β),, interleukin-10 (IL-10) and interferon-γ (IFN-γ) was determined by reverse transcription and real time polymerase chain reaction. Results - Using an in vitro endotoxin challenge method PGG-Glucan altered the production of TNFα in a dose-dependent manner. PGG-Glucan had no effect upon the ex vivo cytokine mRNA expression of TNFα, IL-1β, IL-10 or IFN-γ. Conclusions and Relevance - Although mild changes were observed in TNFα production in vitro, it is not likely that PGG-Glucan will have a significant effect upon clinical endotoxemia.
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