Nystatin Modulates Genes in Immunity and Wingless Signaling Pathways in Cow Blood

Abstract

Nystatin is an antifungal agent isolated from bacteria found in the dairy cow environment. It disrupts small platforms in the cell membrane, composed of sphingolipids and cholesterol known as lipid rafts. Pathogen recognition receptors (PRR) may be embedded in these lipid rafts. This study was conducted to evaluate the in vitro effects of the lipid raft inhibitor Nystatin, on the expression of genes in the innate and adaptive immunity and wnt signaling pathway in cow peripheral blood. Blood collected from four adult female Holstein-Friesian cows (n=4) was treated with 100ng/mL of Nystatin in vitro. Samples treated with Phosphate Buffer Saline served as control. Total protein concentration and prostaglandin E2in plasma were determined. Total RNA was isolated from cells and was used for cDNA synthesis. The effect of Nystatin on the expression of 84 genes on the cow Wingless signaling pathway and human innate and adaptive immunity arrays were assessed in cow blood using real-time PCR. Fold change in transcript abundance was calculated using Livak’s method. Nystatin was found to modulate transcription and translation of genes involved in homeostasis and immunity in cow blood. It also increased the concentration of total plasma protein and PGE2in cow blood and may thus have had a pro-inflammatory effect. This study provides evidence for the association between lipid raft inhibition and alterations in the wingless signaling pathway in ruminant blood. Furthermore, the results presented may inform antifungal drug design and use in cows.