ABSTRACT
Interferon tau, a 23 kDa trophoblast derived protein diffuses out from the uterus into the circulation and leads to the expression of IFNτ stimulated genes viz. ISG15 and OAS1 in blood neutrophils. The IFNτ pathway is species as well as tissue specific. To unsnarl the IFNτ downstream signaling pathway, the blood neutrophils were incubated simultaneously with 10 ng/ml of recombinant bovine interferon tau and the inhibitors of JAK2 (AG490), JAK3 (CP690550), p38 (SB202190), PI3K/Akt (LY294002), and MAPK/Erk (U0126) at specific doses for 4-hr duration. The IFNτ pathway was determined through real-time gene expression of ISG15 and OAS1; immunocytochemistry of ISG15; and Western blotting of ISG15, OAS1, pJAK3 and PI3K. The ISG15 and OAS1 expression decreased significantly (p < 0.001) in the presence of pJAK3 and PI3K inhibitors as compared to a positive control where only interferon tau was used. Immunocytochemistry revealed an attenuated ISG15 response while stimulating blood neutrophils with pJAK3 inhibitor (CP690550) and PI3K inhibitor (LY294002). Similarly, Western blot analysis of neutrophil protein fraction showed weak signals of ISG15, OAS1, pJAK3 and PI3K in the presence of pJAK3 and PI3K inhibitors. The expression profile, immunocytochemistry and western blot analysis revealed a JAK3 and PI3K mediated interferon-tau stimulated gene expression in blood neutrophils.
Subject(s)
Gene Expression Regulation/drug effects , Interferon Type I/pharmacology , Janus Kinase 3/metabolism , Neutrophils/drug effects , Phosphatidylinositol 3-Kinase/metabolism , Pregnancy Proteins/pharmacology , 2',5'-Oligoadenylate Synthetase/genetics , 2',5'-Oligoadenylate Synthetase/metabolism , Animals , Cattle , Cells, Cultured , Female , Neutrophil Activation/drug effects , Neutrophils/enzymology , Phagocytosis/drug effects , Phosphorylation , Recombinant Proteins/pharmacology , Signal Transduction/drug effects , Ubiquitins/genetics , Ubiquitins/metabolismABSTRACT
Serum electrolytes, minerals, blood biochemical and plasma enzymes were studied in heat exposed buffalo heifers supplemented with yeast powder in feed to alleviate the adverse effects of heat stress. Eighteen healthy Murrah buffalo heifers (270-280) kg were divided into three groups. Animals of group I were kept in shed and served as control, while group II and group III were exposed in a psychrometric chamber at 40°C for 4 hrs daily for 16 days continuously. The animals in group III were also supplemented with yeast powder at 10 g per animal per day. The serum sodium, potassium and chloride levels decreased significantly (P < 0.05) in group II and their levels tended to be maintained in group III. Serum calcium, magnesium, phosphorus, glucose and total cholesterol concentration decreased (P < 0.05) significantly whereas total protein increased in group II due to heat stress. The levels of these minerals tended to be normal in group III. The plasma concentration of SGOT and SGPT increased significantly (P < 0.05), whereas the concentration of plasma alkaline phosphatase and acid phosphatase decreased due to thermal exposure. The levels of these enzymes tended to be normal in group III. The results indicated significant deviations in blood biochemical due to thermal stress and their levels tended to be normal in yeast supplemented group of buffalo heifers.
