Bovine Endometrial Cells Mount Innate Immune Response to the Intracellular Ligands CL097 and Poly(dA:dT) Indicating Roles against Uterine Viruses  

作　　者：Chike F. Oguejiofor Zhangrui Cheng Ali A. Fouladi-Nashta D. Claire Wathes 

机构地区：[1]Department of Production and Population Health, Royal Veterinary College, North Mymms, UK [2]Department of Comparative Biomedical Sciences, Royal Veterinary College, North Mymms, UK

出　　处：《Open Journal of Animal Sciences》2017年第2期110-126,共17页动物科学期刊（英文）

摘　　要：Uterine infection and endometritis cause infertility and economic losses in the cattle industry. The innate immune response of the endometrium is critical in the elimination of pathogenic organisms that invade the uterus in postpartum cows. This study investigated the response of bovine endometrium to synthetic intracellular ligands which activate innate immunity by stimulating similar receptors to those used to recognise the presence of some viruses. Mixed primary epithelial and stromal cell cultures were treated with 5 μg/ml of CL097 (a TLR7/8 ligand) or 2 μg/ml of poly(dA:dT) (a DNA analogue) for either 6 h or 24 h. Cellular responses were assessed by the mRNA expression of 18 immune-related genes and 3 endogenous reference genes by conventional PCR followed by qRT-PCR from four replicate experiments. Bovine endometrial cells expressed the cytosolic pattern recognition receptors (PRRs) DDX58 (RIG-I), IFIH1 (MDA5) and LRRFIP1 which act as intracellular nucleic acid sensors. Neither ligand altered the expression of the extra-cytosolic pattern recognition receptors (PRRs) TLR3, TLR4, TLR7 or TLR8 whereas poly(dA:dT) treatment increased the expression of IFIH1 and DDX58. Treated cells also responded to CL097 or poly(dA:dT) with a differential up-regulation of genes involved in innate immune response including type I interferon/antiviral response (MX1, IFNAR1), antimicrobial activity (MUC1, SLPI) and cytokine activity (TNF, IL1B, IL8). Bovine endometrial cells therefore express both cytosolic and extra-cytosolic intracellular PRRs and are able to mount an innate immune response upon stimulation with intracellular ligands. This suggests an important role for these cells in the defence against viruses that may be present in the uterus in postpartum cows.Uterine infection and endometritis cause infertility and economic losses in the cattle industry. The innate immune response of the endometrium is critical in the elimination of pathogenic organisms that invade the uterus in postpartum cows. This study investigated the response of bovine endometrium to synthetic intracellular ligands which activate innate immunity by stimulating similar receptors to those used to recognise the presence of some viruses. Mixed primary epithelial and stromal cell cultures were treated with 5 μg/ml of CL097 (a TLR7/8 ligand) or 2 μg/ml of poly(dA:dT) (a DNA analogue) for either 6 h or 24 h. Cellular responses were assessed by the mRNA expression of 18 immune-related genes and 3 endogenous reference genes by conventional PCR followed by qRT-PCR from four replicate experiments. Bovine endometrial cells expressed the cytosolic pattern recognition receptors (PRRs) DDX58 (RIG-I), IFIH1 (MDA5) and LRRFIP1 which act as intracellular nucleic acid sensors. Neither ligand altered the expression of the extra-cytosolic pattern recognition receptors (PRRs) TLR3, TLR4, TLR7 or TLR8 whereas poly(dA:dT) treatment increased the expression of IFIH1 and DDX58. Treated cells also responded to CL097 or poly(dA:dT) with a differential up-regulation of genes involved in innate immune response including type I interferon/antiviral response (MX1, IFNAR1), antimicrobial activity (MUC1, SLPI) and cytokine activity (TNF, IL1B, IL8). Bovine endometrial cells therefore express both cytosolic and extra-cytosolic intracellular PRRs and are able to mount an innate immune response upon stimulation with intracellular ligands. This suggests an important role for these cells in the defence against viruses that may be present in the uterus in postpartum cows.

关 键 词：ENDOMETRIUM INNATE Immunity INTRACELLULAR LIGANDS Gene Expression 

分 类 号：R73[医药卫生—肿瘤]