Buffalo gut microbes may affect the host Th2 responses during Fasciola gigantica infection via SCFAs metabolism and TLR signaling pathway
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PROVIDER: PRJEB30859 | ENA |
REPOSITORIES: ENA
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Project description:BackgroundFasciola gigantica, the tropical liver fluke, infects buffaloes in Asian and African countries and causes significant economic losses and poses public health threat in these countries. However, little is known of the transcriptional response of buffaloes to infection with F. gigantica. The objective of the present study was to perform the first transcriptomic analysis of buffalo liver infected by F. gigantica. Understanding the mechanisms that underpin F. gigantica infection in buffaloes will contribute to our ability to control this parasite.MethodsWe challenged buffaloes with 500 viable F. gigantica metacercariae and collected liver samples through a time course at 3, 42 and 70 days post-infection (dpi). Then, we performed gene expression analysis on liver samples using RNA sequencing (RNA-Seq) Illumina technology and confirmed the RNA-Seq data by quantitative RT-PCR analysis.ResultsTotals of 496, 880 and 441 differentially expressed transcripts were identified in the infected livers at 3, 42 and 70 dpi, respectively. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that transcriptional changes in the liver of infected buffaloes evolve over the course of infection. The predominant response of buffaloes to infection was mediated by certain pathways, such as MHC antigen processing and presentation, Toll-like receptor 4 (TLR4), transforming growth factor beta (TGF-β), and the cytochrome P450. Hepatic drug metabolizing enzymes and bile secretion were also affected.ConclusionsFasciola gigantica can induce statistically significant and biologically plausible differences in the hepatic gene expression of infected buffaloes. These findings provide new insights into the response of buffaloes to F. gigantica over the course of infection, which may be useful in determining pathways that can modulate host-parasite interaction and thus potentially important for clearance of the parasite.
| S-EPMC5286860 | biostudies-literature
Project description:Widespread Fasciola gigantica infection in buffaloes has caused great economic losses in buffalo farming. Studies on F. gigantica excretory and secretory products (FgESP) have highlighted their importance in F. gigantica parasitism and their potential in vaccine development. Identifying FgESP components involved in F. gigantica-buffalo interactions during different periods is important for developing effective strategies against fasciolosis. Buffaloes were assigned to non-infection (n = 3, as control group) and infection (n = 3) groups. The infection group was orally administrated 250 metacercariae. Sera were collected at 3, 10, and 16 weeks post-infection (wpi) for the non-infection group and at 0 (pre-infection), 1, 3, 6, 8, 10, 13, and 16 wpi for the infection group. FgESP components interacting with sera from the non-infection and infection groups assay were pulled down by co-IP and identified using LC-MS/MS. Interacting FgESP components in infection group were subjected to Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathway and gene ontology (GO) functional annotation to infer their potential functions. Proteins of FgESP components identified in the non-infection group at 3, 10, and 16 wpi accounted for 80.5%, 84.3%, and 82.1% of all proteins identified in these 3 time points, respectively, indicating surroundings did not affect buffalo immune response during maintenance. 490 proteins were identified in the infection group, of which 87 were consistently identified at 7 time points. Following GO analysis showed that most of these 87 proteins were in biological processes, while KEGG analysis showed they mainly functioned in metabolism and cellular processing, some of which were thought to functions throughout the infection process. The numbers of specific interactors identified for each week were 1 (n = 12), 3 (n = 5), 6 (n = 8), 8 (n = 15), 10 (n = 23), 13 (n = 22), and 16 (n = 14) wpi, some of which were thought to functions in specific infection precess. This study screened the antigenic targets in FgESP during a dense time course over a long period. These findings may enhance the understanding of molecular F. gigantica-buffalo interactions and help identify new potential vaccine and drug target candidates.
2022-12-12 | PXD038582 | Pride
Project description:BackgroundExcretory/secretory products (ESPs) released by parasites influence the development and functions of host dendritic cells (DCs). However, little is known about changes of DNA (hydroxy)methylation on DC development during Fasciola gigantica infection. The present study aimed to investigate whether F. gigantica ESPs (FgESPs) affects the development and functions of buffalo DCs through altering the DNA (hydroxy)methylation of DCs.MethodsBuffalo DCs were prepared from peripheral blood mononuclear cells (PBMCs) and characterized using scanning and transmission electron microscopy (SEM/TEM) and quantitative reverse transcriptional PCR (qRT-RCR). DCs were treated with 200 μg/ml of FgESPs in vitro, following DNA extraction. The DNA methylome and hydroxymethylome were profiled based on (hydroxy)methylated DNA immunoprecipitation sequencing [(h)MeDIP-Seq] and bioinformatics analyses. qRT-RCR was also performed to assess the gene transcription levels of interest.ResultsFgESPs markedly suppressed DC maturation evidenced by morphological changes and downregulated gene expression of CD1a and MHC II. Totals of 5432 and 360 genes with significant changes in the 5-methylcytosine (5-mC) and the 5-hydroxymethylcytosine (5-hmC) levels, respectively, were identified in buffalo DCs in response to FgESPs challenge. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that these differentially expressed genes were highly enriched in pathways associated with immune response. Some cancer-related pathways were also indicated. There were 111 genes demonstrating changes in both 5-mC and 5-hmC levels, 12 of which were interconnected and enriched in 12 pathways. The transcription of hypermethylated genes TLR2, TLR4 and IL-12B were downregulated or in a decreasing trend, while the mRNA level of high-hydroxymethylated TNF gene was upregulated in buffalo DCs post-exposure to FgESPs in vitro.ConclusionsTo our knowledge, the present study provides for the first time a unique genome-wide profile of DNA (hydroxy)methylation for DCs that interact with FgESPs, and suggests a possible mechanism of FgESPs in suppressing DC maturation and functions that are involved in TLR signaling.
| S-EPMC6647289 | biostudies-literature
Project description:BackgroundFascioliasis is a zoonotic parasitic disease caused by Fasciola hepatica and Fasciola gigantica, which poses a serious threat to global public health and livestock farming. Fasciola gigantica secretes and excretes various components to manipulate the immune response, thereby enhancing its invasion, migration, and survival in vivo. However, the roles of specific components in immune modulation, such as asparagine endopeptidase, remain unknown.MethodsThe transcriptional abundance of members of the asparagine endopeptidase family (also known as the legumain family) from F. gigantica was analyzed. Two highly transcribed asparagine endopeptidases in metacercariae, juveniles and adults were cloned, and their recombinant proteins-recombinant F. gigantica legumain (rFgLGMN-1) and (rFgLGMN-2)-were expressed in prokaryotic expression system. Their regulatory effects on buffalo peripheral blood mononuclear cells (PBMCs), including proliferation, migration, total nitric oxide (NO) production, cytokine secretion, and phagocytosis were explored in vitro.ResultsTen members of the legumain family were detected in F. gigantica, among of which FgLGMN-1 and FgLGMN-2 exhibited high transcription levels in juveniles and adults. The isolation of sequences indicated that FgLGMN-1 encodes 409 amino acids, while FgLGMN-2 encodes 403 amino acids. Both recombinant FgLGMN-1 (rFgLGMN-1) and rFgLGMN-2 were recognized by serum from buffaloes infected with F. gigantica. Both rFgLGMN-1 and rFgLGMN-2 inhibited the proliferation of PBMCs, and rFgLGMN-1 also inhibited the migration of PBMCs. While rFgLGMN-1 increased the production of total NO, rFgLGMN-2 decreased NO production. Both rFgLGMN-1 and rFgLGMN-2 increased the transcription of the cytokines interleukin-10 and transforming growth factor β. The effect of rFgLGMN-1 and rFgLGMN-2 on the phagocytosis of PBMCs varied depending on their concentrations.ConclusionsrFgLGMN-1 and rFgLGMN-2 modulate several cellular and immunological functions of PBMCs, and exhibited distinct regulatory effects on these in vitro, which indicated that they may play roles in immune modulation and facilitate fluke development. However, due to uncertainties associated with in vitro experiments, further studies are necessary to elucidate the precise functions of these legumains.
| S-EPMC11656647 | biostudies-literature
Project description:Fasciola gigantica infects buffalo in Asian and African countries and causes significant economic losses. As a kind of zoonotic parasite disease, human infection has been reported and make fascioliasis patients suffer heavy abdominal pain, anemia and workforce loss. Fasciola can modulate host immune responses to survive from immune attack and some mechanisms have been reported with its antigen protein. Protein, as the chief actors within the cell, presents the dynamic changes of body with its specific changes and serves as unexceptionable biomarkers in many disease-detections. But there is not any study about serum proteomic for diagnostic target seeking in buffalo after infection with F. gigantica. Herein, Isobaric Tags for Relative and Absolute Quantitation (iTRAQ) and Parallel Reaction Monitoring (PRM) technologies were used in the present study to reveal the dynamic changes of proteins in buffalo serum after infected with F. gigantica. Six proteins were found significantly regulated in all three groups which reminds us that they could be used as new diagnostic biomarkers in buffalo throughout F. gigantica infection. All these are new findings in buffalo after infection with F. gigantica and could provide a new insight into the potential possibility in this parasite clearance.
2019-11-12 | PXD011576 | Pride
Project description:This SuperSeries is composed of the SubSeries listed below.
2019-09-25 | GSE125736 | GEO
Project description:Purpose: To investigate whether FgESP affects the hydroxylmethylation level of DCs Methods: we have profiled the DNA hydroxymethylome by using hydroxymethylated DNA immunoprecipitation sequencing (h)MeDIP-Seq in the current study Results: DNA hydroxymethylation profiles revealed significant changes in the 5-hydroxymethylcytosine (5hmC) levels in 360 genes when DCs were treated with FgESPs. These differentially expressed genes were highly enriched in pathways associated with immune function. Conclusion: For the first time, our study provides a genome-wide mapping of DNA hydroxylmethylation for DCs that interacted with FgESPs.
2019-09-25 | GSE125735 | GEO
Project description:Purpose: To investigate whether FgESP affects the (hydroxyl)methylation level of DCs Methods: we have profiled the DNA methylome by using methylated DNA immunoprecipitation sequencing (MeDIP-Seq) in the current study Results: DNA methylation and hydroxymethylation profiles revealed significant changes in the 5-methylcytosine (5-mC) levels in the promoter regions of 5432 genes and significant changes in the 5-hydroxymethylcytosine (5hmC) levels in 360 genes when DCs were treated with FgESPs. These differentially expressed genes were highly enriched in pathways associated with immune function. The characterizations of DCs were assessed using scanning and transmission electron microscopy (SEM/TEM) and quantitative reverse transcriptional PCR (qRT-PCR). Our results demonstrated that FgESPs could markedly suppress DC maturation by down-regulating the expression of membrane phenotypic markers, such as CD1a, MHC II. The functional enrichment analysis found hypermethylation of genes, especially in the NF-κB signaling pathway, and transcription was unchanged or low for the TLR2, TLR4, and IL-12 genes in DCs. Thus, we suggest a possible mechanism of FgESPs in suppressing TLR signaling pathway-dependent DC maturation and function. Moreover, there were 111 genes demonstrating changes in both 5-mC and 5-hmC levels, and 12 of which were enriched and interconnected in pathways associated with the inflammatory response. It is interesting to note that certain cancer-related pathways were indicated in our analysis, such as ‘pathways in cancer’. Conclusion: For the first time, our study provides a genome-wide mapping of DNA (hydroxyl)methylation for DCs that interacted with FgESPs. Our study also suggests a possible immunoregulatory mechanism of FgESPs that acts by increasing DNA methylation levels on genes and pathways associated with TLR signaling.
2019-09-25 | GSE125734 | GEO
Project description:Background: Intramuscular fat (IMF) content is an important index for beef quality. However, the genetics of IMF deposition is complex and still largely unclear, especially in buffalo. To identify miRNAs with potential regulatory role in lipid accumulated in muscle, we performed small RNA sequencing and identified miRNAs expressed in the longissimus dorsi muscle and back fat of Chinese buffalo, which provided vital information for further identification of miRNAs with potential regulatory role in the lipid accumulated in muscle. Results: Three small RNA libraries were constructed. A total of 32762032 raw reads were obtained from adipose groups, respectively. After filtering the adaptor and low quality reads, 32054381 clean reads were retained. In total, 623 miRNAs were identified.
2021-12-31 | GSE171146 | GEO
Project description:Background: Intramuscular fat (IMF) content is an important index for beef quality. However, the genetics of IMF deposition is complex and still largely unclear, especially in buffalo. To identify miRNAs with potential regulatory role in lipid accumulated in muscle, we performed small RNA sequencing and identified miRNAs expressed in the longissimus dorsi muscle and back fat of Chinese buffalo, which provided vital information for further identification of miRNAs with potential regulatory role in the lipid accumulated in muscle. Results: Six small RNA libraries were constructed. A total of 66,128,645 and 70,974,347 raw reads were obtained from muscle and adipose groups, respectively. After filtering the adaptor and low quality reads, 60,765,257 and 67,327,095 clean reads were retained. In total, 721 miRNAs were identified.
2021-08-27 | GSE119064 | GEO