Modulation of C. parvum Interaction with Bovine PMN

Bovine PMN were pre-treated with the mentioned inhibitors for 30 min and then co-cultured with C. parvum sporozoites (1:3 PMN/sporozoite ratio, 3 h, 37 °C) with the purpose of blocking the ATP purinergic receptor P2X1, MCT1, MCT2, and glycolysis under hyperoxia 21% O₂ = oxygen conditions widely used in most C. parvum-related studies) and physioxia (5% O₂) conditions, simulating physiological oxygen pressure of small intestine in vivo as follows [4 (link),12 (link),54 (link),59 (link)]: AR-C 155858 (1 μM, MCT1- and MCT2-inhibitors, Tocris Bioscience, Wiesbaden, Germany), AR-C 141990 (1 μM, MCT1 inhibitor, Tocris Bioscience, Wiesbaden, Germany), NF449 (100 μM, purinergic receptor P2X1 antagonist, Tocris Bioscience, Wiesbaden, Germany), oligomycin A (5 μM, inhibitor of mitochondrial respiration, Sigma-Aldrich, Darmstadt, Germany) and 2-DG (2 mM; antagonist of glycolysis, Sigma-Aldrich, Darmstadt, Germany), were used in this study [37 (link),44 (link),60 (link),61 (link)].