Evaluation of Pgg-Glucan, a Novel Immunomodulator, in in Vitro and Ex Vivo Models of Equine Endotoxemia
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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 Î¼g/ml) and LPS (100 ng/ml) plus PGG-Glucan (0.1, 1.0, 10 and 100 Î¼gg/ml). Supernatants were collected at 0, 6 and 12 hours and assayed for tumor necrosis factor Â£\ (TNFÂ£\) 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 TNFÎ±, interleukin-1Î² (IL-1Î²),, interleukin-10 (IL-10) and interferon-Î³ (IFN-Î³) 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 TNFÎ± in a dose-dependent manner. PGG-Glucan had no effect upon the ex vivo cytokine mRNA expression of TNFÎ±, IL-1Î², IL-10 or IFN-Î³.
Conclusions and Relevance - Although mild changes were observed in TNFÎ± production in vitro, it is not likely that PGG-Glucan will have a significant effect upon clinical endotoxemia.
- Masters Theses