ABSTRACT: Transcriptomic and microRNA profiling of equine and viral genes in peripheral blood mononuclear cells in horses during equine herpesvirus 1 infection
Project description:To investigate pre-existing mucosal immunity in equine herpesvirus type 1 (EHV-1) immune and non-immune horses during experimental viral challenge. RNAseq was performed on nasal mucosal swab samples from immune and non-immune horses from pre (d-2), early (d1pi and d3pi), mid (d8pi and d10pi), and late (d18pi) infection.
Project description:Equine hepacivirus (EqHV) is the closest genetic relative of hepatitis C virus (HCV) and shares features of genome organization, hepatotropism, persistent infection, and the ability to cause liver disease. As such, EqHV studies are important both in order to understand equine liver disease, and as an outbred animal model for HCV pathogenesis and immune responses. Here, we characterize the natural history and immune response to EqHV infection. Seven horses were experimentally inoculated with EqHV, monitored for 6 months, and challenge inoculated with the same, and subsequently a divergent EqHV inoculum. Clearance was the primary outcome (6 of 7) and was associated with subclinical hepatitis characterized by lymphocytic infiltrate and individual hepatocyte necrosis. Seroconversion was delayed and antibody titers waned slowly. Resolving horses developed non-sterilizing immunity resulting in short duration of infection upon challenge. Unlike those observed in acutely HCV-infected patients, peripheral blood mononuclear cell responses in horses were minimal, although EqHV specific T-cells were identified. In contrast, an interferon stimulated gene signature was detected in the liver during EqHV infection, which is similar to acute HCV in humans. EqHV, similarly to HCV, is stimulated by direct binding of the liver-specific microRNA, miR-122. Surprisingly, we found that EqHV infection sequesters enough miR-122 to functionally affect gene regulation in the liver. This RNA-based mechanism thus could have consequences for pathology. Conclusion: EqHV infection in horses typically has an acute resolving course, and the immune response attenuates subsequent infections lasting for at least a year. This could have important implications to achieve the first goal of an HCV vaccine; to prevent chronicity while accepting acute resolving infection after challenge.
2020-09-30 | GSE158753 | GEO
Project description:MicroRNA sequencing in equine peripheral blood mononuclear cells
| PRJNA352827 | ENA
Project description:Identification of Host and Viral Factors Associated with the Development of Equine Herpesvirus Myeloencephalopathy (EHM) by Transcriptomic and microRNA Analysis of Peripheral Blood Mononuclear Cells from Horses
Project description:Equine hepacivirus (EqHV) is phylogenetically the closest relative of hepatitis C virus (HCV) and shares genome organization, hepatotropism, transient or persistent infection outcome, and the ability to cause hepatitis. Thus, EqHV studies are important to understand equine liver disease, and further as an outbred surrogate animal model for HCV pathogenesis and protective immune responses. Here, we aimed to characterize the course of EqHV infection and associated protective immune responses. Approach & Results: Seven horses were experimentally inoculated with EqHV, monitored for 6 months, and rechallenged with the same, and subsequently a heterologous EqHV. Clearance was the primary outcome (6 of 7) and was associated with subclinical hepatitis characterized by lymphocytic infiltrate and individual hepatocyte necrosis. Seroconversion was delayed and antibody titers waned slowly. Clearance of primary infection conferred non-sterilizing immunity resulting in shortened duration of viremia after rechallenge. Peripheral blood mononuclear cell responses in horses were minimal, although EqHV specific T cells were identified. Additionally, an interferon stimulated gene signature was detected in the liver during EqHV infection, similar to acute HCV in humans. EqHV, as HCV, is stimulated by direct binding of the liver-specific microRNA, miR-122. Interestingly, we found that EqHV infection sequesters enough miR-122 to functionally affect gene regulation in the liver. This RNA-based mechanism thus could have consequences for pathology. Conclusions: EqHV infection in horses typically has an acute resolving course, and the protective immune response lasts for at least a year and broadly attenuates subsequent infections. This could have important implications to achieve the primary goal of an HCV vaccine; to prevent chronicity while accepting acute resolving infection after virus exposure.
Project description:Capacity of exercise and performance is the most valuable in the horses. They have been selected for strength, speed, and indurance trait. Athletic pheno types are influenced markedly by environment, management, and training. However, it has long been accepted that there are underlying genetic factors. To determine altered mRNA expression in circulating leukocytes of horses induced by exercise. Healthy neutered male warmblood horses were subjected to indoor exercise (trotting with alternative cantering for 6o minutes). Peripheral blood was collected from the jugular vein before and after the exercise, and subsequently buffy coat leukocytes were isolated by centrifugation. Total RNAs was isolated. Cyanine 3-labeled cRNA (complementary RNA) was generated from Agilentâs Low RNA Input Linear Amplification kit with 500 ng total RNA. Labeled cRNA was applied microarray (Agilent technologies, 8x60K) using Agilentâs Gene Expression Hybridization Kit. The present study revealed a subset of mRNAs in equine peripheral blood leukocytes affected by exercise, providing background information for genes associated with exercise in warm-blood horses. Three healthy, gelding warmblood horses between 9 and 17 yr were selected. 6 samples were collected containing 3 samples before exercise and 3 samples after exercise
Project description:Capacity of exercise and performance is the most valuable in the horses. They have been selected for strength, speed, and indurance trait. Athletic pheno types are influenced markedly by environment, management, and training. However, it has long been accepted that there are underlying genetic factors. To determine altered mRNA expression in circulating leukocytes of horses induced by exercise. Healthy neutered male warmblood horses were subjected to indoor exercise (trotting with alternative cantering for 6o minutes). Peripheral blood was collected from the jugular vein before and after the exercise, and subsequently buffy coat leukocytes were isolated by centrifugation. Total RNAs was isolated. Cyanine 3-labeled cRNA (complementary RNA) was generated from Agilent’s Low RNA Input Linear Amplification kit with 500 ng total RNA. Labeled cRNA was applied microarray (Agilent technologies, 8x60K) using Agilent’s Gene Expression Hybridization Kit. The present study revealed a subset of mRNAs in equine peripheral blood leukocytes affected by exercise, providing background information for genes associated with exercise in warm-blood horses.
Project description:Equine Papillomavirus Type 2 (EcPV2) appears to be a causal factor for the development of genital especially penile squamous cell carcinomas (SCC) and as such have an important clinical impact on horses. However, the pathomechanisms associated with this cancer transformation are not known, yet. To analyze the host’s and viral transcriptome in EcPV2 affected horses, tissue samples were collected from horses with EcPV2-positive genital lesions as well as from healthy EcPV2-negative horses. Expression levels of host and viral genes were evaluated by RNA-Seq.