Project description:Background: Fasciola hepatica infection still remains one of the helminthic neglected tropical diseases (NTDs). It has a huge worldwide distribution, affecting mainly cattle and, sometimes, human beings. In addition to data reported about the immunological response induced by helminthic infections and that induced by Fasciola hepatica, little is known about the gene expression profile in its organ target, the liver, which is where adult worms are established and live for long periods of time, causing its characteristic pathology. In the present work, we study both the early and late gene expression profiles in the livers of mice infected with Fasciola hepatica metacercariae using a microarray-based methodology. Methodology: A total of 9 female-6-week-old BALB/c mice (Charles River Laboratories, Barcelona, Spain) weighing 20 to 35 g were used for the experiments. Two groups of BALB/c mice were orally infected with seven F. hepatica metacercariae, and the other group remained untreated and served as a control. Mice were humanely euthanized and necropsied for liver recovery, histological assessment of hepatic damage, RNA isolation, microarray design and gene expression analysis on the day of infection (t0), seven days post-infection (t7) and twenty-one days post-infection (t21). Results: We found that Fasciola hepatica infection induces the differential expression of 128 genes in the liver in the early stage of infection and 308 genes in the late stage, and most of them are up-regulated. The Ingenuity Pathway Analysis revealed significant changes in the pathways related to metabolism, biosynthesis and signaling as well as genes implicated in inducing liver-toxicity, injury and death. Conclusion: The present study provides us insights at the molecular level about the underlying mechanisms used by Fasciola hepatica, leading to liver damage and its subsequent pathophysiology. The expression pattern obtained here could also be used to explain the lack of association between infection with F. hepatica and cholangiocarcinoma. However, more studies should be performed to confirm this hypothesis. We used three experimental groups each containing 3 mice. Group 1 remains untreated and served as control. Group 2 was infected with Fasciola hepatica metacercariae on day 0 and humanely necropsied at 7 days post-infection. Group 3 was infected with Fasciola hepatica metacercariae on day 0 and humanely necropsied at 21 days post- infection. At the time of necropsy, liver of each mice were removed and the RNA was isolated. We compared the gene expression profile in the liver of mice infected with Fasciola hepatica.
Project description:Helminths, or worms, are multicellular parasites that can live for many years within their vertebrate hosts. Of prime importance is the regulation of the host immune cell signalling pathways to prevent the parasite’s elimination before they can produce their off-spring in the form of eggs. Fasciola hepatica, a global worm parasite of humans and their livestock, regulates host innate immune responses within hours of infection. Host macrophages, essential to the first-line defence mechanisms, are quickly restricted in their ability to initiate a classic protective pro-inflammatory immune response. To determine the role of both host miRNAs and parasite-derived miRNAs in this outcome to infection with F. hepatica, peritoneal macrophages were harvested from the peritoneal cavity of BALB/c mice at various timepoints during infection and subject to RNASeq. Sequenced Ago2 extracted from peritoneal macrophages of fasciola infected mice
Project description:Background: Fasciola hepatica infection still remains one of the helminthic neglected tropical diseases (NTDs). It has a huge worldwide distribution, affecting mainly cattle and, sometimes, human beings. In addition to data reported about the immunological response induced by helminthic infections and that induced by Fasciola hepatica, little is known about the gene expression profile in its organ target, the liver, which is where adult worms are established and live for long periods of time, causing its characteristic pathology. In the present work, we study both the early and late gene expression profiles in the livers of mice infected with Fasciola hepatica metacercariae using a microarray-based methodology. Methodology: A total of 9 female-6-week-old BALB/c mice (Charles River Laboratories, Barcelona, Spain) weighing 20 to 35 g were used for the experiments. Two groups of BALB/c mice were orally infected with seven F. hepatica metacercariae, and the other group remained untreated and served as a control. Mice were humanely euthanized and necropsied for liver recovery, histological assessment of hepatic damage, RNA isolation, microarray design and gene expression analysis on the day of infection (t0), seven days post-infection (t7) and twenty-one days post-infection (t21). Results: We found that Fasciola hepatica infection induces the differential expression of 128 genes in the liver in the early stage of infection and 308 genes in the late stage, and most of them are up-regulated. The Ingenuity Pathway Analysis revealed significant changes in the pathways related to metabolism, biosynthesis and signaling as well as genes implicated in inducing liver-toxicity, injury and death. Conclusion: The present study provides us insights at the molecular level about the underlying mechanisms used by Fasciola hepatica, leading to liver damage and its subsequent pathophysiology. The expression pattern obtained here could also be used to explain the lack of association between infection with F. hepatica and cholangiocarcinoma. However, more studies should be performed to confirm this hypothesis.
Project description:F. hepatica is a parasite causing losses of great importance in animal production. It is also considered a zoonosis of growing importance. During the infection the parasite releases a number of antigens (called Excretory Secretory Products - ESP) facilitating the parasites survival. A number of ESP components have immunomodulatory properties. To determine if ES from various F. hepatica isolates have different immunomodulatory abilities “BOMA” cells were stimulated with ES from laboratory Weybridge (FhWey-ES) and wild (FhWild-ES) Fasciola hepatica isolates.
Project description:Fasciola hepatica represents a socioeconomically important species of parasitic fluke which causes significant economic losses in the livestock industry. During an indirect life cycle, immature eggs are passed out of the host in the stool and embyonate in the external environment until the release of miracidium which invades a freshwater snail - intermediate host. By utilization of RNA-seq technique (Illumina NextSeq 500), differentially expressed genes were analysed in F. hepatica eggs during the maturation process in three time points: 0 days (freshly laid eggs by adult F. hepatica individuals) and after incubation for 5 and 10 days.
Project description:F. hepatica is a parasite causing losses of great importance in animal production. It is also considered a zoonosis of growing importance. During the infection the parasite releases a number of antigens (called Excretory Secretory Products - ESP) facilitating the parasites survival. A number of ESP components have immunomodulatory properties. To determine if ES from various F. hepatica isolates have different immunomodulatory abilities LPS activated “BOMA” cells were stimulated with ES of laboratory Weybridge (FhWey-ES) and wild (FhWild-ES) Fasciola hepatica isolates.
Project description:Liver fluke (Fasciola hepatica) infection is both a welfare and productivity issue in sheep farming. Death can result from acute infection, and deaths are becoming more frequent as anthelmintic resistance increases. New control strategies are desirable, and vaccination is a good option. Previous studies have shown an experimental vaccine based on a F. hepatica protein, cathepsin L1, (rmFhCL1) may be a viable aid to control liver fluke in cattle/sheep, however efficacy is variable. A trial was conducted on sheep immunized with rmFhCL1 following infection with F. hepatica to understand the immune response changes induced by vaccinination at a molecular level . Peripheral blood mononuclear cells (PBMCs) were isolated at four different time points for RNA-Seq analysis. Genes differentially expressed between vaccinated and control animals were identified. Their functional roles were studied using in silico methods.