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Xing-quan Zhu - One of the best experts on this subject based on the ideXlab platform.

  • Global serum proteomic changes in water buffaloes infected with Fasciola Gigantica
    Parasites & vectors, 2019
    Co-Authors: Fu-kai Zhang, Hany M. Elsheikha, Zhao-an Sheng, Wei-yu Zhang, Wen-bin Zheng, Xing-quan Zhu
    Abstract:

    The liver fluke Fasciola Gigantica modulates several signaling pathways in infected buffaloes to facilitate its survival and establishment of persistent infection. In response to the parasite invasion, buffaloes activate innate and adaptive immune responses to counter the parasite infection. To detect new proteins that might be involved in the interaction between F. Gigantica and the buffaloes, and that also might serve as biomarkers for fasciolosis, we used proteomic techniques to study the serum proteome of buffaloes during F. Gigantica infection. Here, we used an isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomic approach to identify serum proteins that are differentially expressed in infected buffaloes compared to uninfected control buffaloes. Additionally, we applied a parallel reaction monitoring (PRM) assay to validate specific proteins identified by the iTRAQ method. A total of 313, 459 and 399 proteins were identified at 3, 42 and 70 days post-infection, respectively; of these 92, 93 and 138 were differentially abundant proteins. Some of the identified differentially abundant proteins, including complement factor H related 5, complement component C6, complement component C7, amine oxidase, plasma serine protease inhibitor and lysozyme, are known to be involved in complement system activation, blood coagulation, platelet activation, lymphocyte’s adhesion and lysozyme hydrolysis. Analysis of data for all three time points after infection identified six significantly upregulated proteins in infected serum that separated infected and uninfected buffaloes into distinct clusters. Further PRM analysis confirmed the expression of five proteins, namely MHC class I antigen, Beta-2-microglobulin, NID2 protein, Fetuin-B and Fibrinogen gamma-B chain. These findings provide novel insights into the serum proteomics signature of buffaloes during F. Gigantica infection.

  • serum levels of cytokines in water buffaloes experimentally infected with Fasciola Gigantica
    Veterinary Parasitology, 2017
    Co-Authors: Fu-kai Zhang, Hany M. Elsheikha, Zhao-an Sheng, Xiaoxuan Zhang, Weiyi Huang, Aijiang Guo, Junling Hou, Miaomiao Sun, Xing-quan Zhu
    Abstract:

    Fasciola Gigantica infection in water buffaloes causes significant economic losses especially 27 in developing countries. Although modulation of the host immune response by cytokine 28 neutralization or vaccination is a promising approach to control infection with this parasite, our 29 understanding of cytokine's dynamic during F. Gigantica infection is limited. To address this, 30 we quantified the levels of serum cytokines produced in water buffaloes following experimental 31 infection with F. Gigantica. Five buffaloes were infected via oral gavage with 500 viable F. 32 Gigantica metacercariae and blood samples were collected from buffaloes one week before 33 infection and for 13 consecutive weeks thereafter. The levels of 10 cytokines in serum samples 34 were simultaneously determined using ELISA. F. Gigantica failed to elicit the production of 35 various pro-inflammatory cytokines, including interleukin-1β (IL-1β), IL-2, IL-6, IL-12, and 36 IFN-γ. On the other hand, evidence of a Th2 type response was detected, but only early in the 37 course of parasite colonization and included modest increase in the levels of IL-10 and IL-13. 38 The results also revealed suppression of the immune responses as a feature of chronic F. 39 Gigantica infection in buffaloes. Taken together, F. Gigantica seems to elicit a modest Th2 40 response at early stage of infection in order to downregulate harmful Th1- and Th17-type 41 inflammatory responses in experimentally infected buffaloes. The full extent of anti-F. 42 Gigantica immune response and its relation to pathogenesis requires further study.

  • transcriptomic responses of water buffalo liver to infection with the digenetic fluke Fasciola Gigantica
    Parasites & Vectors, 2017
    Co-Authors: Fu-kai Zhang, Hany M. Elsheikha, Zhao-an Sheng, Wen-bin Zheng, Xiaoxuan Zhang, Weiyi Huang, Aijiang Guo, Xing-quan Zhu
    Abstract:

    Background Fasciola 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. Methods We 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. Results Totals 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. Conclusions Fasciola 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.

  • transcriptomic responses of water buffalo liver to infection with the digenetic fluke Fasciola Gigantica
    Parasites & Vectors, 2017
    Co-Authors: Fu-kai Zhang, Hany M. Elsheikha, Zhao-an Sheng, Wen-bin Zheng, Xiaoxuan Zhang, Weiyi Huang, Aijiang Guo, Xing-quan Zhu
    Abstract:

    Fasciola 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. We 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. Totals 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. Fasciola 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.

Prasert Sobhon - One of the best experts on this subject based on the ideXlab platform.

  • cytosolic superoxide dismutase can provide protection against Fasciola Gigantica
    Acta Tropica, 2016
    Co-Authors: Wipaphorn Jaikua, Pornanan Kueakhai, Prasert Sobhon, Kulathida Chaithirayanon, Rataya Tanomrat, Sirima Wongwairot, Suda Riengrojpitak, Narin Changklungmoa
    Abstract:

    Superoxide dismutases (SOD), antioxidant metallo-enzymes, are a part of the first line of defense in the trematode parasites which act as the chief scavengers for reactive oxygen species (ROS). A recombinant Fasciola Gigantica cytosolic SOD (FgSOD) was expressed in Escherichia coli BL21 (DE3) and used for immunizing rabbits to obtain polyclonal antibodies (anti-rFgSOD). This rabbit anti-rFgSOD reacted with the native FgSOD at a molecular weight of 17.5kDa. The FgSOD protein was expressed at high level in parenchyma, caecal epithelium and egg of the parasite. The rFgSOD reacted with antisera from rabbits infected with F. Gigantica metacercariae collected at 2, 5, and 7 weeks after infection, and reacted with sera of infected mice. Anti-rFgSOD exhibited cross reactivity with the other parasites' antigens, including Eurytrema pancreaticum, Cotylophoron cotylophorum, Fischoederius cobboldi, Gastrothylax crumenifer, Paramphistomum cervi, and Setaria labiato papillosa. A vaccination was performed in imprinting control region (ICR) mice by subcutaneous injection with 50μg of rFgSOD combined with Freund's adjuvant. At 2 weeks after the second boost, mice were infected with 15 metacercariae by oral route. IgG1 and IgG2a in the immune sera were determined to indicate Th2 and Th1 immune responses. It was found that the parasite burden was reduced by 45%, and both IgG1 and IgG2a levels showed correlation with the numbers of worm recoveries.

  • immunodiagnosis of Fasciola Gigantica infection using monoclonal antibody based sandwich elisa and immunochromatographic assay for detection of circulating cathepsin l1 protease
    PLOS ONE, 2016
    Co-Authors: Panat Anuracpreeda, Runglawan Chawengkirttikul, Prasert Sobhon
    Abstract:

    Background Tropical fasciolosis caused by Fasciola Gigantica infection is one of the major diseases infecting ruminants in the tropical regions of Africa and Asia including Thailand. Parasitological diagnosis of fasciolosis is often unreliable and possesses low sensitivity. Therefore, the detection of circulating parasite antigens is thought to be a better alternative for diagnosis of fasciolosis, as it reflects the real parasite burden. Methods In this study, we have produced a monoclonal antibody (MoAb) against recombinant F. Gigantica cathepsin L1 (rFgCatL1), and developed both sandwich enzyme-linked immunosorbent assay (sandwich ELISA) and immunochromatographic (IC) test for rapid detection of circulating cathepsin L1 protease (CatL1) in the sera from mice experimentally and cattle naturally infected with Fasciola Gigantica. MoAb 4E3 and biotinylated rabbit anti-recombinant CatL1 antibody were selected due to their high reactivities and specificities. Results The lower detection limits of sandwich ELISA and IC test were 3 pg/ml and 0.256 ng/ml, respectively. Sandwich ELISA and IC test could detect F. Gigantica infection from day 1 to 35 post infection. In experimental mice, the sensitivity, specificity and accuracy were 95%, 100% and 98.6% (for sandwich ELISA), and 93%, 100% and 98.2% (for IC test), while in natural cattle they were 98.3%, 100% and 99.5% (for sandwich ELISA), and 96.7%, 100% and 99.1% (for IC test). Conclusions These two assay methods showed high efficiencies and precisions for diagnosis of fasciolosis by F. Gigantica.

  • immunization with recombinant leucine aminopeptidase showed protection against Fasciola Gigantica in mice
    Parasitology Research, 2013
    Co-Authors: Narin Changklungmoa, Pathanin Chantree, Pannigan Chaichanasak, Pornanan Kueakhai, Tadashi Itagaki, Kulathida Chaithirayanon, Suda Riengrojpitak, Veerawat Sansri, Narin Preyavichyapugdee, Prasert Sobhon
    Abstract:

    Leucine aminopeptidase (LAP) is expressed in all stages of Fasciola Gigantica and, hence, is considered as a potential vaccine candidate. In this study, we have tested a vaccine potential of LAP and the types of immune responses it elicited in vaccinated mice. Recombinant F. Gigantica leucine aminopeptidase (rFgLAP) was expressed in Escherichia coli, BL21 (DE3). The imprinting control region mice subcutaneously immunized with 50 μg of rFgLAP combined with Freund's adjuvant (n = 10) exhibited a significant reduction in worm recoveries when compared with non-immunized and Freund's adjuvant controls at 60.8 and 64.3%, respectively, and both T helper (Th)1 and Th2 humoral immune responses were elicited in the hosts as reflected by the levels of IgG1 and IgG2a, with Th2 predominating. The levels of IgG1- and IgG2a-specific antibodies to rFgLAP were inversely and significantly correlated with the numbers of worm recoveries. The rFgLAP-vaccinated mice showed significantly reduced levels of serum glutamic oxaloacetic transaminase and serum glutamic pyruvic transaminase and liver damage. These indicated that rFgLAP has a potential as a vaccine candidate against F. Gigantica, whose efficacy will be studied further in economic animals including cattle, sheep, and goat.

  • production and characterization of a monoclonal antibody against recombinant saposin like protein 2 of Fasciola Gigantica
    Acta Tropica, 2013
    Co-Authors: Pornanan Kueakhai, Narin Changklungmoa, Sineenart Songkoomkrong, Kulathida Chaithirayanon, Suda Riengrojpitak, Prasert Sobhon
    Abstract:

    A monoclonal antibody (MoAb) against recombinant Fasciola Gigantica saposin-like protein 2 (rFgSAP-2) was produced by hybridoma technique using spleen cells from BALB/c mice immunized with rFgSAP-2. This MoAb is an IgG1, κ light chain isotype. By immunoblotting and indirect ELISA, the MoAb reacted specifically with rFgSAP-2, the natural FgSAP-2 at 10kDa in whole body (WB) and excretory-secretory (ES) fractions of F. Gigantica. It did not cross react with antigens in WB fractions from other parasites, including Opisthorchis viverrini, Schistosoma mansoni which are human parasites, Haemonchus placei, Setaria labiato-papillosa, Eurytrema pancreaticum, Cotylophoron cotylophorum, Fischoederius cobboldi, Gigantocotyle explanatum, Gastrothylax crumenifer, and Paramphistomum cervi which are ruminant parasites. By immunohistochemistry, the FgSAP-2 protein was localized only in the cytoplasm of caecal epithelial cells of 4-week-old juvenile and adult stages, but not in metacercariae, newly excysted juvenile (NEJ), 2- and 3-week-old juveniles. This finding indicated that FgSAP-2 is an abundantly expressed parasite protein that is released into the ES, hence SAP-2 and its MoAb may be used for immunodiagnosis of ruminant and human fasciolosis.

  • characterization and expression of cathepsin b2 in Fasciola Gigantica
    Experimental Parasitology, 2012
    Co-Authors: Pathanin Chantree, Manussabhorn Phatsara, Krai Meemon, Chaitip Wanichanon, Prasert Sobhon
    Abstract:

    Abstract Fasciola Gigantica cathepsin B belongs to a family of cysteine proteases which is involved in invasion of host tissues. In this study, the recombinant cathepsin B2 (rFgCatB2), synthesized in Pichia pastoris, showed enzymatic activity on a fluorometric substrate Z-Phe-Arg-AMC and gelatin. Furthermore, this recombinant enzyme could degrade IgG and type I collagen. Mouse antiserum against rFgCatB2 reacted with the native FgCatB2 in whole body (WB) extracts of metacercariae (MET), newly excysted juveniles (NEJ) and 2 week-old juveniles, but not in 3, 4 week-old juveniles and adult flukes. Immunolocalization showed the presence of cathepsin B2 only in the caecal epithelium of MET, NEJ and 2 week-old juveniles. Co-localization of FgCatB2 and a prominent antigen of NEJ, FgCatB3, revealed that these proteins were expressed at the same regions in the caecal epithelium. Anti-rFgCatB2 showed no cross reaction with the other parasites’ antigens by Western blotting. These findings suggest that CatB2 is expressed only in early stages of the parasite and may be involved in digestion of host connective tissues and evasion of the host immune system during their penetration and migration. Thus, CatB2 could be considered as an immunodiagnostic and vaccine candidate for fasciolosis.

Fu-kai Zhang - One of the best experts on this subject based on the ideXlab platform.

  • Global serum proteomic changes in water buffaloes infected with Fasciola Gigantica
    Parasites & vectors, 2019
    Co-Authors: Fu-kai Zhang, Hany M. Elsheikha, Zhao-an Sheng, Wei-yu Zhang, Wen-bin Zheng, Xing-quan Zhu
    Abstract:

    The liver fluke Fasciola Gigantica modulates several signaling pathways in infected buffaloes to facilitate its survival and establishment of persistent infection. In response to the parasite invasion, buffaloes activate innate and adaptive immune responses to counter the parasite infection. To detect new proteins that might be involved in the interaction between F. Gigantica and the buffaloes, and that also might serve as biomarkers for fasciolosis, we used proteomic techniques to study the serum proteome of buffaloes during F. Gigantica infection. Here, we used an isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomic approach to identify serum proteins that are differentially expressed in infected buffaloes compared to uninfected control buffaloes. Additionally, we applied a parallel reaction monitoring (PRM) assay to validate specific proteins identified by the iTRAQ method. A total of 313, 459 and 399 proteins were identified at 3, 42 and 70 days post-infection, respectively; of these 92, 93 and 138 were differentially abundant proteins. Some of the identified differentially abundant proteins, including complement factor H related 5, complement component C6, complement component C7, amine oxidase, plasma serine protease inhibitor and lysozyme, are known to be involved in complement system activation, blood coagulation, platelet activation, lymphocyte’s adhesion and lysozyme hydrolysis. Analysis of data for all three time points after infection identified six significantly upregulated proteins in infected serum that separated infected and uninfected buffaloes into distinct clusters. Further PRM analysis confirmed the expression of five proteins, namely MHC class I antigen, Beta-2-microglobulin, NID2 protein, Fetuin-B and Fibrinogen gamma-B chain. These findings provide novel insights into the serum proteomics signature of buffaloes during F. Gigantica infection.

  • serum levels of cytokines in water buffaloes experimentally infected with Fasciola Gigantica
    Veterinary Parasitology, 2017
    Co-Authors: Fu-kai Zhang, Hany M. Elsheikha, Zhao-an Sheng, Xiaoxuan Zhang, Weiyi Huang, Aijiang Guo, Junling Hou, Miaomiao Sun, Xing-quan Zhu
    Abstract:

    Fasciola Gigantica infection in water buffaloes causes significant economic losses especially 27 in developing countries. Although modulation of the host immune response by cytokine 28 neutralization or vaccination is a promising approach to control infection with this parasite, our 29 understanding of cytokine's dynamic during F. Gigantica infection is limited. To address this, 30 we quantified the levels of serum cytokines produced in water buffaloes following experimental 31 infection with F. Gigantica. Five buffaloes were infected via oral gavage with 500 viable F. 32 Gigantica metacercariae and blood samples were collected from buffaloes one week before 33 infection and for 13 consecutive weeks thereafter. The levels of 10 cytokines in serum samples 34 were simultaneously determined using ELISA. F. Gigantica failed to elicit the production of 35 various pro-inflammatory cytokines, including interleukin-1β (IL-1β), IL-2, IL-6, IL-12, and 36 IFN-γ. On the other hand, evidence of a Th2 type response was detected, but only early in the 37 course of parasite colonization and included modest increase in the levels of IL-10 and IL-13. 38 The results also revealed suppression of the immune responses as a feature of chronic F. 39 Gigantica infection in buffaloes. Taken together, F. Gigantica seems to elicit a modest Th2 40 response at early stage of infection in order to downregulate harmful Th1- and Th17-type 41 inflammatory responses in experimentally infected buffaloes. The full extent of anti-F. 42 Gigantica immune response and its relation to pathogenesis requires further study.

  • transcriptomic responses of water buffalo liver to infection with the digenetic fluke Fasciola Gigantica
    Parasites & Vectors, 2017
    Co-Authors: Fu-kai Zhang, Hany M. Elsheikha, Zhao-an Sheng, Wen-bin Zheng, Xiaoxuan Zhang, Weiyi Huang, Aijiang Guo, Xing-quan Zhu
    Abstract:

    Background Fasciola 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. Methods We 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. Results Totals 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. Conclusions Fasciola 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.

  • transcriptomic responses of water buffalo liver to infection with the digenetic fluke Fasciola Gigantica
    Parasites & Vectors, 2017
    Co-Authors: Fu-kai Zhang, Hany M. Elsheikha, Zhao-an Sheng, Wen-bin Zheng, Xiaoxuan Zhang, Weiyi Huang, Aijiang Guo, Xing-quan Zhu
    Abstract:

    Fasciola 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. We 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. Totals 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. Fasciola 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.

Hany M. Elsheikha - One of the best experts on this subject based on the ideXlab platform.

  • Global serum proteomic changes in water buffaloes infected with Fasciola Gigantica
    Parasites & vectors, 2019
    Co-Authors: Fu-kai Zhang, Hany M. Elsheikha, Zhao-an Sheng, Wei-yu Zhang, Wen-bin Zheng, Xing-quan Zhu
    Abstract:

    The liver fluke Fasciola Gigantica modulates several signaling pathways in infected buffaloes to facilitate its survival and establishment of persistent infection. In response to the parasite invasion, buffaloes activate innate and adaptive immune responses to counter the parasite infection. To detect new proteins that might be involved in the interaction between F. Gigantica and the buffaloes, and that also might serve as biomarkers for fasciolosis, we used proteomic techniques to study the serum proteome of buffaloes during F. Gigantica infection. Here, we used an isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomic approach to identify serum proteins that are differentially expressed in infected buffaloes compared to uninfected control buffaloes. Additionally, we applied a parallel reaction monitoring (PRM) assay to validate specific proteins identified by the iTRAQ method. A total of 313, 459 and 399 proteins were identified at 3, 42 and 70 days post-infection, respectively; of these 92, 93 and 138 were differentially abundant proteins. Some of the identified differentially abundant proteins, including complement factor H related 5, complement component C6, complement component C7, amine oxidase, plasma serine protease inhibitor and lysozyme, are known to be involved in complement system activation, blood coagulation, platelet activation, lymphocyte’s adhesion and lysozyme hydrolysis. Analysis of data for all three time points after infection identified six significantly upregulated proteins in infected serum that separated infected and uninfected buffaloes into distinct clusters. Further PRM analysis confirmed the expression of five proteins, namely MHC class I antigen, Beta-2-microglobulin, NID2 protein, Fetuin-B and Fibrinogen gamma-B chain. These findings provide novel insights into the serum proteomics signature of buffaloes during F. Gigantica infection.

  • serum levels of cytokines in water buffaloes experimentally infected with Fasciola Gigantica
    Veterinary Parasitology, 2017
    Co-Authors: Fu-kai Zhang, Hany M. Elsheikha, Zhao-an Sheng, Xiaoxuan Zhang, Weiyi Huang, Aijiang Guo, Junling Hou, Miaomiao Sun, Xing-quan Zhu
    Abstract:

    Fasciola Gigantica infection in water buffaloes causes significant economic losses especially 27 in developing countries. Although modulation of the host immune response by cytokine 28 neutralization or vaccination is a promising approach to control infection with this parasite, our 29 understanding of cytokine's dynamic during F. Gigantica infection is limited. To address this, 30 we quantified the levels of serum cytokines produced in water buffaloes following experimental 31 infection with F. Gigantica. Five buffaloes were infected via oral gavage with 500 viable F. 32 Gigantica metacercariae and blood samples were collected from buffaloes one week before 33 infection and for 13 consecutive weeks thereafter. The levels of 10 cytokines in serum samples 34 were simultaneously determined using ELISA. F. Gigantica failed to elicit the production of 35 various pro-inflammatory cytokines, including interleukin-1β (IL-1β), IL-2, IL-6, IL-12, and 36 IFN-γ. On the other hand, evidence of a Th2 type response was detected, but only early in the 37 course of parasite colonization and included modest increase in the levels of IL-10 and IL-13. 38 The results also revealed suppression of the immune responses as a feature of chronic F. 39 Gigantica infection in buffaloes. Taken together, F. Gigantica seems to elicit a modest Th2 40 response at early stage of infection in order to downregulate harmful Th1- and Th17-type 41 inflammatory responses in experimentally infected buffaloes. The full extent of anti-F. 42 Gigantica immune response and its relation to pathogenesis requires further study.

  • transcriptomic responses of water buffalo liver to infection with the digenetic fluke Fasciola Gigantica
    Parasites & Vectors, 2017
    Co-Authors: Fu-kai Zhang, Hany M. Elsheikha, Zhao-an Sheng, Wen-bin Zheng, Xiaoxuan Zhang, Weiyi Huang, Aijiang Guo, Xing-quan Zhu
    Abstract:

    Background Fasciola 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. Methods We 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. Results Totals 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. Conclusions Fasciola 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.

  • transcriptomic responses of water buffalo liver to infection with the digenetic fluke Fasciola Gigantica
    Parasites & Vectors, 2017
    Co-Authors: Fu-kai Zhang, Hany M. Elsheikha, Zhao-an Sheng, Wen-bin Zheng, Xiaoxuan Zhang, Weiyi Huang, Aijiang Guo, Xing-quan Zhu
    Abstract:

    Fasciola 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. We 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. Totals 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. Fasciola 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.

  • rapid identification and differentiation of Fasciola hepatica and Fasciola Gigantica by a loop mediated isothermal amplification lamp assay
    Veterinary Parasitology, 2010
    Co-Authors: Hany M. Elsheikha, Sungjong Hong, J X Chen, S H Chen, X Q Cai, M X Chen, X Q Zhu
    Abstract:

    Abstract The present study developed and validated a species-specific loop-mediated isothermal amplification (LAMP) assay for the rapid detection and discrimination of Fasciola hepatica and Fasciola Gigantica. The LAMP assay is inexpensive, easy to perform and shows rapid reaction, wherein the amplification can be obtained in 45 min under isothermal conditions of 61 °C or 62 °C by employing a set of four species-specific primer mixtures and results can be checked through naked-eye visualization. The optimal assay conditions with no cross-reaction with other closely related trematodes (Clonorchis sinensis, Opisthorchis viverrini, Orientobilharzia turkestanicum and Schistosoma japonicum) as well as within the two Fasciola species were established. The assay was validated by examining F. Gigantica DNA in the intermediate host snails and in faecal samples. The results indicated that the LAMP assay is approximately 104 times more sensitive than the conventional specific PCR assays. These findings indicate that this Fasciola species-specific LAMP assay may have a potential clinical application for detection and differentiation of Fasciola species, especially in endemic countries.

D Sriveny - One of the best experts on this subject based on the ideXlab platform.

  • immuno diagnosis of bubaline fasciolosis with Fasciola Gigantica cathepsin l and recombinant cathepsin l 1 d proteases
    Acta Tropica, 2006
    Co-Authors: O K Raina, S C Yadav, D Sriveny, S C Gupta
    Abstract:

    Abstract Fasciola Gigantica cathepsin-L cysteine proteinase and recombinant cathepsin L 1-D were assessed for their potential in the immuno-diagnosis of F. Gigantica infection in buffaloes. A diagnostic ELISA, based on these two antigens, was developed to detect antibodies against F. Gigantica in water buffaloes. Sensitivity of the ELISA was assessed using sera from buffaloes experimentally or naturally infected with F. Gigantica from F. Gigantica endemic areas and its specificity by probing the sera of the host from F. Gigantica non-endemic area. Our earlier studies under experimental setting showed 100% sensitivity of cathepsin-L ELISA in the diagnosis of fasciolosis in buffaloes, with the earliest detection of infection at 4 weeks post-infection. However, under field situation of natural F. Gigantica infection , this sensitivity declined to 97.1% but specificity of the test remained 100%. Cross-reactivity of the antigen was checked with Schistosoma indicum , S. spindale , Paramphistomum epiclitum , Gastrothylax spp., Gigantocotyle explanatum , hydatid and Strongyloides papilossus in the bubaline host, naturally infected with these helminths. F. Gigantica cathepsin-L and the recombinant cathepsin L-1D does not cross-react with these helminth parasites in natural mono or mixed infection of the host. The present ELISA contributes a relatively sensitive and reliable tool for the early serodiagnosis of bubaline fasciolosis.

  • cathepsin l cysteine proteinase in the diagnosis of bovine Fasciola Gigantica infection
    Veterinary Parasitology, 2006
    Co-Authors: D Sriveny, S C Yadav, O K Raina, D Chandra, A K Jayraw, Mithilesh Singh, R Velusamy, B P Singh
    Abstract:

    Cathepsin L cysteine proteinase from Fasciola Gigantica was evaluated for its potential in the early prepatent detection of this helminth infection in bovine calves. Five cross-bred bovine calves were experimentally infected with 400 metacercariae/calf and evaluated for anti-cathepsin L antibody response. F. Gigantica infection in these calves could be detected 4 weeks post-infection using an ELISA, dipstick ELISA and Western blotting with 100% sensitivity. The antigen was also used to detect F. Gigantica field infection in cattle, by screening 256 sera of these animals by an ELISA, which demonstrated an overall infection rate of 26.95%. Preliminary studies showed that F. Gigantica cathepsin L cysteine proteinase does not cross-react with Paramphistomum epiclitum, Gigantocotyle explanatum and hydatid cyst antigens. However, extensive studies on the cross-reactivity of this antigen with related helminth parasites of cattle and buffaloes are required, before this antigen can be considered suitable for immuno-diagnosis of fasciolosis in these ruminants.

  • Fasciola Gigantica cathepsin l cysteine proteinase in the detection of early experimental fasciolosis in ruminants
    Parasitology Research, 2005
    Co-Authors: S C Yadav, Mohini Saini, O K Raina, P A Nambi, K Jadav, D Sriveny
    Abstract:

    Cathepsin-L cysteine proteinase was purified from Fasciola Gigantica regurgitant by two-step alcoholic fractionation, followed by ion-exchange chromatography. The purification strategy was evolved to eliminate other contaminating proteins co-precipitating with the purified proteinase during alcoholic fractionation. The enzyme was stable on long-term storage at –20°C rendering it more suitable for field diagnostic use. The purified cathepsin-L cysteine proteinase was assayed for detection of F. Gigantica experimental infection in sheep and buffaloes and could detect infection, as early as 4 weeks post-infection by ELISA, Western blotting and Dipstick ELISA. The 28-kDa cathepsin-L cysteine proteinase seems a promising antigen for the diagnosis of tropical fasciolosis in domestic animals.

  • Fasciola Gigantica cathepsin l cysteine proteinase in the detection of early experimental fasciolosis in ruminants
    Parasitology Research, 2005
    Co-Authors: S C Yadav, Mohini Saini, O K Raina, P A Nambi, K Jadav, D Sriveny
    Abstract:

    Cathepsin-L cysteine proteinase was purified from Fasciola Gigantica regurgitant by two-step alcoholic fractionation, followed by ion-exchange chromatography. The purification strategy was evolved to eliminate other contaminating proteins co-precipitating with the purified proteinase during alcoholic fractionation. The enzyme was stable on long-term storage at -20 degrees C rendering it more suitable for field diagnostic use. The purified cathepsin-L cysteine proteinase was assayed for detection of F. Gigantica experimental infection in sheep and buffaloes and could detect infection, as early as 4 weeks post-infection by ELISA, Western blotting and Dipstick ELISA. The 28-kDa cathepsin-L cysteine proteinase seems a promising antigen for the diagnosis of tropical fasciolosis in domestic animals.