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Anja Joachim - One of the best experts on this subject based on the ideXlab platform.
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the parasitic nematode Oesophagostomum dentatum synthesizes unusual glycosaminoglycan like o glycans
Glycobiology, 2018Co-Authors: Jorick Vanbeselaere, Anja Joachim, Katharina Paschinger, Iain B H WilsonAbstract:: O-glycosylation is probably one of the most varied sets of post-translational modifications across all organisms, but amongst the most refractory to analyze. In animals, O-xylosylation of serine residues represents the first stage in the synthesis of glycosaminoglycans, whose repeat regions are generally analyzed as fragments resulting from enzymatic or chemical degradation, whereas their core regions can be isolated by β-elimination or endo-β-xylosidase digestion. In the present study, we show that hydrazinolysis can be employed for release of glycosaminoglycan-type oligosaccharides from nematodes prior to fluorescent labeling with 2-aminopyridine. While various [HexNAcHexA]nGal2Xyl oligosaccharides were isolated from the model organism Caenorhabditis elegans, more unusual glycosaminoglycan-type glycans were found to be present in the porcine parasite Oesophagostomum dentatum. In this case, as judged by MS/MS before and after hydrofluoric acid or β-galactosidase digestion, core sequences with extra galactose and phosphorylcholine residues were detected as [(±PC)HexNAcHexA]n(±PC)Galβ3-(±Galβ4)Galβ4Xyl. Thus, hydrazinolysis and fluorescent labeling can be combined to analyze unique forms of O-xylosylation, including new examples of zwitterionic glycan modifications.
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gender and developmental specific n glycomes of the porcine parasite Oesophagostomum dentatum
Biochimica et Biophysica Acta, 2017Co-Authors: Carmen Jimenezcastells, Bärbel Ruttkowski, Anja Joachim, Jorick Vanbeselaere, Sonja Kohlhuber, Katharina PaschingerAbstract:Abstract Background The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. However, in contrast to the non-parasitic C. elegans, the parasitic O. dentatum is an obligate sexual organism, which makes both a gender and developmental glycomic comparison possible. Methods Different enzymatic and chemical methods were used to release N-glycans from male and female O. dentatum as well as from L3 and L4 larvae. Glycans were analysed by MALDI-TOF MS after either 2D-HPLC (normal then reversed phase) or fused core RP-HPLC. Results Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. Seemingly, simple methylated paucimannosidic structures can be considered ‘male’, while methylation of fucosylated glycans was more pronounced in females. On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2–4PC2–4 motifs not observed in our previous studies on other nematodes. Conclusion The interspecies tendency of class V nematodes to share most, but not all, N-glycans applies also to O. dentatum; furthermore, we establish, for the first time in a parasitic nematode, that glycomes vary upon development and sexual differentiation. General significance Unusual methylated, core fucosylated and phosphorylcholine-containing N-glycans vary between stages and genders in a parasitic nematode.
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Oesophagostomum dentatum Extract Modulates T Cell- Dependent Immune Responses to Bystander Antigens and Prevents the Development of Allergy in Mice
2016Co-Authors: Irma Schabussova, Anja Joachim, Onisa Ul-haq, Elisabeth Hoflehner, Angelika Wagner, Gerhard Loupal, Rick M. Maizels, Ursula WiedermannAbstract:One third of the human population is currently infected by one or more species of parasitic helminths. Certain helminths establish long-term chronic infections resulting in a modulation of the host’s immune system with attenuated responsiveness to ‘‘bystander’ ’ antigens such as allergens or vaccines. In this study we investigated whether parasite-derived products suppress the development of allergic inflammation in a mouse model. We show that extract derived from adult male Oesophagostomum dentatum (eMOD) induced Th2 and regulatory responses in BALB/c mice. Stimulation of bone marrow-derived dendritic cells induced production of regulatory cytokines IL-10 and TGF-beta. In a mouse model of birch pollen allergy, co-administration of eMOD with sensitizing allergen Bet v 1 markedly reduced the production of allergen-specific antibodies in serum as well as IgE-dependent basophil degranulation. Furthermore, eMOD prevented the development of airway inflammation, as demonstrated by attenuation of bronchoalveolar lavages eosinophil influx, peribronchial inflammatory infiltrate, and mucus secretion in lungs and IL-4 and IL-5 levels in lung cell cultures. Reduced secretion of Th2-related cytokines by birch pollen-re-stimulated splenocytes and mesenteric lymph node cells was observed in eMOD-treated/sensitized and challenged mice in comparison to sensitized and challenged controls. The suppressive effects of eMOD were heat-stable. Immunization with model antigens in the presence of eMOD reduced production of antibodies to thymus-dependent but not to thymus-independent antigen, suggesting that suppression of the immun
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recent advances in elucidating nematode moulting prospects of using Oesophagostomum dentatum as a model
Advances in Parasitology, 2016Co-Authors: Martina Ondrovics, Robin B Gasser, Anja JoachimAbstract:There are major gaps in our knowledge of many molecular biological processes that take place during the development of parasitic nematodes, in spite of the fact that understanding such processes could lead to new ways of treating and controlling parasitic diseases via the disruption of one or more biological pathways in the parasites. Progress in genomics, transcriptomics, proteomics and bioinformatics now provides unique opportunities to investigate the molecular basis of key developmental processes in parasitic nematodes. The porcine nodule worm, Oesophagostomum dentatum, represents a large order (Strongylida) of socioeconomically important nematodes, and provides a useful platform for exploring molecular developmental processes, particularly given that this nematode can be grown and maintained in culture in vitro for periods longer than most other nematodes of this order. In this article, we focus on the moulting process (ecdysis) in nematodes; review recent advances in our understanding of molecular aspects of moulting in O. dentatum achieved by using integrated proteomic-bioinformatic tools and discuss key implications and future prospects for research in this area, also with respect to developing new anti-nematode interventions and biotechnological outcomes.
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proteomics elucidates key molecules involved in exsheathment in vitro in Oesophagostomum dentatum
International Journal for Parasitology, 2014Co-Authors: Martina Ondrovics, Robin B Gasser, Katja Silbermayr, Makedonka Mitreva, Neil D. Young, Anja JoachimAbstract:We explored molecules involved in in vitro exsheathment of Oesophagostomum dentatum L3s using a proteomic-transcriptomic-bioinformatic approach. Analysis of L3s before, during and after exsheathment identified 11 proteins that were over-expressed exclusively during exsheathment. These proteins (including key enzymes, heat shock, structural and nematode-specific proteins) were inferred to be involved in development, metabolism, structure, motility and/or host-parasite interactions. Some of these molecules represented homologues linked to entry into and exit from the dauer stage in the free-living nematode Caenorhabditis elegans. The approach established here provides a basis for investigations of ecdysis in other strongylid nematodes.
Robin B Gasser - One of the best experts on this subject based on the ideXlab platform.
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recent advances in elucidating nematode moulting prospects of using Oesophagostomum dentatum as a model
Advances in Parasitology, 2016Co-Authors: Martina Ondrovics, Robin B Gasser, Anja JoachimAbstract:There are major gaps in our knowledge of many molecular biological processes that take place during the development of parasitic nematodes, in spite of the fact that understanding such processes could lead to new ways of treating and controlling parasitic diseases via the disruption of one or more biological pathways in the parasites. Progress in genomics, transcriptomics, proteomics and bioinformatics now provides unique opportunities to investigate the molecular basis of key developmental processes in parasitic nematodes. The porcine nodule worm, Oesophagostomum dentatum, represents a large order (Strongylida) of socioeconomically important nematodes, and provides a useful platform for exploring molecular developmental processes, particularly given that this nematode can be grown and maintained in culture in vitro for periods longer than most other nematodes of this order. In this article, we focus on the moulting process (ecdysis) in nematodes; review recent advances in our understanding of molecular aspects of moulting in O. dentatum achieved by using integrated proteomic-bioinformatic tools and discuss key implications and future prospects for research in this area, also with respect to developing new anti-nematode interventions and biotechnological outcomes.
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proteomics elucidates key molecules involved in exsheathment in vitro in Oesophagostomum dentatum
International Journal for Parasitology, 2014Co-Authors: Martina Ondrovics, Robin B Gasser, Katja Silbermayr, Makedonka Mitreva, Neil D. Young, Anja JoachimAbstract:We explored molecules involved in in vitro exsheathment of Oesophagostomum dentatum L3s using a proteomic-transcriptomic-bioinformatic approach. Analysis of L3s before, during and after exsheathment identified 11 proteins that were over-expressed exclusively during exsheathment. These proteins (including key enzymes, heat shock, structural and nematode-specific proteins) were inferred to be involved in development, metabolism, structure, motility and/or host-parasite interactions. Some of these molecules represented homologues linked to entry into and exit from the dauer stage in the free-living nematode Caenorhabditis elegans. The approach established here provides a basis for investigations of ecdysis in other strongylid nematodes.
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Proteomic Analysis of Oesophagostomum dentatum (Nematoda) during Larval Transition, and the Effects of Hydrolase Inhibitors on Development
PloS one, 2013Co-Authors: Martina Ondrovics, Robin B Gasser, Katja Silbermayr, Makedonka Mitreva, Neil D. Young, Ebrahim Razzazi-fazeli, Anja JoachimAbstract:In this study, in vitro drug testing was combined with proteomic and bioinformatic analyses to identify and characterize proteins involved in larval development of Oesophagostomum dentatum, an economically important parasitic nematode. Four hydrolase inhibitors ο-phenanthroline, sodium fluoride, iodoacetamide and 1,2-epoxy-3-(pnitrophenoxy)-propane (EPNP) significantly inhibited (≥90%) larval development. Comparison of the proteomic profiles of the development-inhibited larvae with those of uninhibited control larvae using two-dimensional gel electrophoresis, and subsequent MALDI-TOF mass spectrometric analysis identified a down-regulation of 12 proteins inferred to be involved in various larval developmental processes, including post-embryonic development and growth. Furthermore, three proteins (i.e. intermediate filament protein B, tropomyosin and peptidyl-prolyl cis-trans isomerase) inferred to be involved in the moulting process were down-regulated in moulting- and development-inhibited O. dentatum larvae. This first proteomic map of O. dentatum larvae provides insights in the protein profile of larval development in this parasitic nematode, and significantly improves our understanding of the fundamental biology of its development. The results and the approach used might assist in developing new interventions against parasitic nematodes by blocking or disrupting their key biological pathways.
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Oesophagostomum dentatum potential as a model for genomic studies of strongylid nematodes with biotechnological prospects
Biotechnology Advances, 2007Co-Authors: Robin B Gasser, Bärbel Ruttkowski, P A Cottee, Alasdair J. Nisbet, Shoba Ranganathan, Anja JoachimAbstract:Abstract There are substantial gaps in the knowledge of the molecular processes of development and reproduction in parasitic nematodes, despite the fact that understanding such processes could lead to novel ways of treating and controlling parasitic diseases, through blocking or disrupting key biological pathways. Biotechnological advances through large-scale sequencing projects, approaches for the analysis of differential gene and protein expression and functional genomics (e.g., double-stranded RNA interference) now provide opportunities to investigate the molecular basis of developmental processes in some parasitic nematodes. The porcine nodule worm, Oesophagostomum dentatum (order Strongylida), may provide a platform for testing the function of genes from this and related nematodes, given that this species can be grown and maintained in culture in vitro for periods longer than other nematodes of the same order. In this article, we review relevant biological, biochemical and molecular biological and genomic information about O. dentatum and propose that the O. dentatum — pig system provides an attractive model for exploring molecular developmental and reproductive processes in strongylid nematodes, leading toward new intervention methods and biotechnological outcomes.
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construction of gender enriched cdna archives for adult Oesophagostomum dentatum by suppressive subtractive hybridization and a microarray analysis of expressed sequence tags
Parasitology, 2006Co-Authors: P A Cottee, Alasdair J. Nisbet, Y Abs G Elosta, T L Webster, Robin B GasserAbstract:: In the present study, we constructed gender-enriched cDNA libraries for the adult stage of the parasitic nematode Oesophagostomum dentatum (order Strongylida) using suppressive-subtractive hybridization (SSH), sequenced clones from the female-library and male-library (480 from each) and conducted bioinformatic and microarray analyses of the expressed sequence tags (ESTs). In total, 873 ESTs (440 male and 433 female) were obtained, achieving a sequencing success of 91%The nucleotide sequences reported in this article (Tables 1-5) have been deposited in the EMBL, GenBank and DDJB databases under the Accession nos. AM157797-AM158083. Microarray analyses of 516 unique ESTs representing both gender-enriched libraries revealed differential hybridization for 391 of them (75.8%). Of these, 220 (56.3%) had significantly greater signal intensities in the female than in the male, and 154 (70%) of these were predicted to have homologues in C. elegans. These homologues were predicted to be involved in key biological processes, including embryonic nutrition, gametogenesis, molecular binding/transport or metabolism, nucleic acid synthesis and function, and signal transduction. Of the 171 ESTs with statistically higher signal intensities in male O. dentatum, 43.8% had homologues in C. elegans. These homologues included major sperm proteins (MSPs) or MSP-like molecules, keratin-like molecules, molecules involved in metabolism, PDZ domain-containing proteins, sugar binding proteins, protein kinases, serine proteases or protease inhibitors, molecules involved in proteolysis and other proteins, such as enzymes and various putative proteins. Of the 287 ESTs (from both gender-enriched cDNA libraries) with no known homologues in C. elegans, 50 (17.4%) had homologues in other nematodes, 8 had homologues in various other organisms and 104 (36.2%) had no homology to any sequence in current gene databases. The present study lays a foundation for the isolation and molecular, biochemical and functional characterization of selected genes from the gender-enriched cDNA archives established for O. dentatum.
Peter Nansen - One of the best experts on this subject based on the ideXlab platform.
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the effect of dietary carbohydrates with different digestibility on the populations of Oesophagostomum dentatum in the intestinal tract of pigs
Parasitology, 2001Co-Authors: Saulius Petkevicius, Peter Nansen, Knud Erik Bach Knudsen, K D MurrellAbstract:: An experiment was undertaken to study the effect of dietary carbohydrates with different digestibility on the populations of Oesophagostomum dentatum in the intestinal tract of pigs. Sixty-four worm-free pigs from a specific pathogen-free farm were randomly divided into 8 equal groups. The animals in 4 groups were assigned to a diet with partially undegradable carbohydrates (diet 1), while the pigs in the 4 remaining groups were given a diet with fermentable carbohydrates (diet 2). Diet 1 was comprised of barley flour, oat husk meal, soybean meal, vitamins and minerals and diet 2 of barley flour, inulin and sugar beet fibre, soybean meal, vitamins and minerals. The pigs in 6 of the groups (n = 48) were inoculated with 6,000 infective larvae of O. dentatum. To determine O. dentatum populations at the early stage of infection, 16 pigs were slaughtered 3 weeks p.i., while the remaining 4 groups continued on the diets for a further 9 weeks after which they were slaughtered. In a diet cross-over experiment 6 weeks after inoculation, 8 pigs changed from diet 1 to diet 2 (diet 1 > diet 2), and 8 pigs from diet 2 to diet 1 (diet 2 > diet 1). The results showed that partially undegradable carbohydrates provided favourable conditions not only for parasite establishment and sustainability, but also for already established O. dentatum infection while, in contrast, the diet composed of highly degradable carbohydrates decreased worm establishment, size and female fecundity. The implications for pastured pigs or pigs fed different complex carbohydrate diets is discussed.
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Use of a migration assay for the separation of adult pyrantel-susceptible and -resistant Oesophagostomum dentatum.
Veterinary parasitology, 2000Co-Authors: Mads Bjelke Petersen, Henrik Bjørn, Jeff Craven, Peter NansenAbstract:Abstract A migration assay was used to separate a pyrantel-susceptible and -resistant isolate of the pig nematode, Oesophagostomum dentatum. The experiment had three steps. In the first step, LD50 values for pyrantel of the two isolates in the assay were established. In the second step, susceptible and resistant worms were mixed in various proportions prior to exposure to a fixed concentration of pyrantel and thereafter assayed. The inhibition of migration showed to be linearly correlated with the proportion of resistant worms in a sample. In step three, this line was used as a standard curve to predict the number of resistant worms in samples from pigs infected with mixed samples of susceptible and resistant larvae.
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The sexually linked Mpi locus is presumably involved in imidothiazole resistance in Oesophagostomum dentatum parasites.
Parasitology research, 2000Co-Authors: V. Šnábel, Peter Nansen, Henrik Bjørn, T. Demeeŵs, Marián Várady, J. ČorbaAbstract:Information about genetic changes during the selection process could indicate mechanisms underlying the spread of resistance to anthelmintic drugs. For clarification of the role of the Mpi locus encoding mannose-phosphate isomerase enzyme in determining resistance, genotyping of Oesophagostomum dentatum strains was performed using an isoelectrofocusing technique. In levamisole- and pyrantel-selected strains the allele associated with resistance has probably been found. Significant values for genetic differentiation between treated and untreated strains of common origin were recorded by Fst indices (θ=0.078; P=0.0008). The specific genomic makeup of a flubendazole-resistant strain, which did not correlate with that of the remaining isolates, might be ascribed to a different action of the anthelmintic or different environmental conditions under which resistance against this drug arose. The absence of heterozygotes in male populations indicated an XX/X0 system of sex determination for the Mpi locus, thus providing a greater potential for the development of resistance. A possible involvement of alleles linked with mannose-phosphate isomerase in alterations of membrane receptors that can be associated with resistance against imidothiazole-based drugs is discussed.
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isolation and characterisation of sex specific transcripts from Oesophagostomum dentatum by rna arbitrarily primed pcr
Molecular and Biochemical Parasitology, 2000Co-Authors: Peter R Boag, C M Christensen, Peter Nansen, Susan E Newton, Nielspeter Hansen, Robin B GasserAbstract:Abstract In light of the lack of molecular data on the sexual differentiation, maturation and interaction of parasitic nematodes of livestock, the present study investigated sex-specific gene expression in the nodule worm, Oesophagostomum dentatum (Strongylida). Using the technique of RNA arbitrarily-primed polymerase chain reaction (RAP-PCR), 31 expressed sequence tags (ESTs) differentially-displayed between the sexes were cloned. Northern blot analysis proved ten ESTs to be expressed exclusively in males (adults and fourth-stage larvae), while two were expressed solely in female stages. None of the ESTs were expressed in infective third-stage larvae. Sequence analysis and subsequent database searches revealed two male-specific ESTs to have significant similarity to Caenorhabditis elegans (predicted) proteins, a protein containing an EGF-like cysteine motif and a serine/threonine phosphatase. Another two male-specific ESTs had similarity to non-nematode sequences. The two female-specific ESTs had similarity to vitellogenin-5 and endonuclease III (predicted) from C. elegans . The remaining ESTs had no similarity to any nucleic acid or protein sequences contained in the databases. The isolation and characterisation of sex-specific ESTs from O. dentatum provides a unique opportunity for studying the reproductive biology of parasitic nematodes at the molecular level, with a view toward novel approaches for parasite control.
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Concurrent Ascaris suum and Oesophagostomum dentatum infections in pigs
Veterinary parasitology, 1999Co-Authors: A B Helwigh, C M Christensen, Andreas Roepstorff, Peter NansenAbstract:Abstract The aim of this study was to examine interactions between Ascaris suum and Oesophagostomum dentatum infections in pigs with regard to population dynamics of the worms such as recovery, location and length; and host reactions such as weight gain, pathological changes in the liver and immune response. Seventy-two helminth-naive pigs were allocated into four groups. Group A was inoculated twice weekly with 10 000 O. dentatum larvae for 8 weeks and subsequently challenge-infected with 1000 A. suum eggs, while Group B was infected with only 1000 A. suum eggs; Group C was inoculated twice weekly with 500 A. suum eggs for 8 weeks and subsequently challenge-infected with 5000 O. dentatum larvae, whereas Group D was given only 5000 O. dentatum larvae. All trickle infections continued until slaughter. Twelve pigs from Group A and B were slaughtered 10 days post challenge infection (p.c.i.) and the remaining 12 pigs from the each of the four groups were slaughtered 28 days p.c.i.. No clinical signs of parasitism were observed. The total worm burdens and the distributions of the challenge infection species were not influenced by previous primary trickle-infections with the heterologous species. Until day 10 p.c.i. the ELISA response between A. suum antigen and sera from the O. dentatum trickle infected pigs (Group A) pigs were significantly higher compared to the uninfected Group B. This was correlated with a significantly higher number of white spots on the liver surface both on Day 10 and 28 p.c.i. in Group A compared to Group B. The mean length of the adult O. dentatum worms was significantly reduced in the A. suum trickle infected group compared to the control group. These results indicate low level of interaction between the two parasite species investigated.
Bärbel Ruttkowski - One of the best experts on this subject based on the ideXlab platform.
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gender and developmental specific n glycomes of the porcine parasite Oesophagostomum dentatum
Biochimica et Biophysica Acta, 2017Co-Authors: Carmen Jimenezcastells, Bärbel Ruttkowski, Anja Joachim, Jorick Vanbeselaere, Sonja Kohlhuber, Katharina PaschingerAbstract:Abstract Background The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. However, in contrast to the non-parasitic C. elegans, the parasitic O. dentatum is an obligate sexual organism, which makes both a gender and developmental glycomic comparison possible. Methods Different enzymatic and chemical methods were used to release N-glycans from male and female O. dentatum as well as from L3 and L4 larvae. Glycans were analysed by MALDI-TOF MS after either 2D-HPLC (normal then reversed phase) or fused core RP-HPLC. Results Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. Seemingly, simple methylated paucimannosidic structures can be considered ‘male’, while methylation of fucosylated glycans was more pronounced in females. On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2–4PC2–4 motifs not observed in our previous studies on other nematodes. Conclusion The interspecies tendency of class V nematodes to share most, but not all, N-glycans applies also to O. dentatum; furthermore, we establish, for the first time in a parasitic nematode, that glycomes vary upon development and sexual differentiation. General significance Unusual methylated, core fucosylated and phosphorylcholine-containing N-glycans vary between stages and genders in a parasitic nematode.
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Oesophagostomum dentatum extract modulates t cell dependent immune responses to bystander antigens and prevents the development of allergy in mice
PLOS ONE, 2013Co-Authors: Irma Schabussova, Bärbel Ruttkowski, Anja Joachim, Elisabeth Hoflehner, Angelika Wagner, Gerhard Loupal, Rick M. Maizels, Onisa Ulhaq, Johnnie Akgun, Ursula WiedermannAbstract:One third of the human population is currently infected by one or more species of parasitic helminths. Certain helminths establish long-term chronic infections resulting in a modulation of the host’s immune system with attenuated responsiveness to “bystander” antigens such as allergens or vaccines. In this study we investigated whether parasite-derived products suppress the development of allergic inflammation in a mouse model. We show that extract derived from adult male Oesophagostomum dentatum (eMOD) induced Th2 and regulatory responses in BALB/c mice. Stimulation of bone marrow-derived dendritic cells induced production of regulatory cytokines IL-10 and TGF-beta. In a mouse model of birch pollen allergy, co-administration of eMOD with sensitizing allergen Bet v 1 markedly reduced the production of allergen-specific antibodies in serum as well as IgE-dependent basophil degranulation. Furthermore, eMOD prevented the development of airway inflammation, as demonstrated by attenuation of bronchoalveolar lavages eosinophil influx, peribronchial inflammatory infiltrate, and mucus secretion in lungs and IL-4 and IL-5 levels in lung cell cultures. Reduced secretion of Th2-related cytokines by birch pollen-re-stimulated splenocytes and mesenteric lymph node cells was observed in eMOD-treated/sensitized and challenged mice in comparison to sensitized and challenged controls. The suppressive effects of eMOD were heat-stable. Immunization with model antigens in the presence of eMOD reduced production of antibodies to thymus-dependent but not to thymus-independent antigen, suggesting that suppression of the immune responses by eMOD was mediated by interference with antigen presenting cell or T helper cell function but did not directly suppress B cell function. In conclusion, we have shown that eMOD possesses immunomodulatory properties and that heat-stable factors in eMOD are responsible for the dramatic suppression of allergic responses in a mouse model of type I allergy. The identification and characterization of parasite-derived immune-modulating molecules might have potential for designing novel prophylactic/therapeutic strategies for immune-mediated diseases.
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transcription profiles for two key gender specific gene families in Oesophagostomum dentatum during development in vivo and in vitro
Infection Genetics and Evolution, 2012Co-Authors: Martina Ondrovics, Bärbel Ruttkowski, A J Nisbet, R B Gasser, Anja JoachimAbstract:Abstract In strongylid roundworms, such as Oesophagostomum dentatum (porcine nodule worm), some sex-specific genes are likely to be associated with parasite maturation, development and reproduction. In this study, an analysis of transcription of the two sex-specific genes ( vit and msp ) encoding vitellogenin and major sperm protein of O. dentatum , respectively, revealed that adult females transcribed vit and adult males msp at high levels, in contrast to immature larval stages and pre-adult worms from in vitro cultures for which no transcription of vit or msp was detected. The analysis showed that neither presence nor absence of the heterologous sex, nor the duration of infection, was central to vit or msp transcription. In small or “virgin” adults, no or only low-level transcription of vit and msp was detectable. We hypothesize that the transcription of the sex-specific genes is linked to endogenous factors, such as size, maturation of the reproductive organs and/or fitness of the worms, and not to exogenous influences. The maturation of worms appears to be linked, to some extent, to the expression of the genes studied herein.
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prostaglandin d2 synthesis in Oesophagostomum dentatum is mediated by cytosolic glutathione s transferase
Experimental Parasitology, 2011Co-Authors: Anja Joachim, Bärbel RuttkowskiAbstract:Abstract Glutathione S -transferases (GSTs) of Oesophagostomum dentatum possess considerable similarity to synthetic prostaglandin D synthase (PGDS), and therefore their ability to convert prostaglandin (PG) H 2 to PGD 2 in vitro was investigated with a commercial Prostaglandin D Synthase Inhibitor Screening Assay Kit. Fractioned homogenates of O. dentatum third-stage larvae only displayed cytosolic but not microsomal GST. Both total larval homogenate and isolated GST could metabolise PGH 2 to PGD 2 , which could be inhibited by the GST inhibitor sulfobromophthalein (SBP) in a dose-dependent manner, whereas reactions to the specific PGDS inhibitor HQL-79 were not dose-dependent. Inhibition of larval development by SBP in vitro was abolished by the addition of PGD 2 but not by PGH 2 , supporting the assumption that GST acts as PGDS and is important for nematode development. Since motility and viability of O. dentatum larvae are reduced in vitro by various inhibitors of eicosanoid metabolism, enzymes of this pathway, including GST, constitute putative intervention targets.
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Oesophagostomum dentatum: effect of glutathione S-transferase (GST) inhibitors on GST activity and larval development.
Experimental Parasitology, 2011Co-Authors: Anja Joachim, Esther Lautscham, Jana Christoffers, Bärbel RuttkowskiAbstract:Abstract Sulphobromophthalein (SBP) inhibits isolated glutathione S -transferase of the porcine nodule worm Oesophagostomum dentatum (Od-GST) and reduces larval development in vitro. In this study possible inhibitory effects of various inhibitors were evaluated in an enzymatic (CDNB) assay with isolated Od-GST and in a larval development assay (LDA). Reversibility was tested in the LDA by removing the inhibitor from culture halfway through the cultivation period. SBP, indomethacin and ethacrynic acid inhibited both enzyme activity and larval development in a dose-dependent and reversible manner. HQL-79 also reduced larval development but had only a minor effect on the isolated enzyme. The phospholipase A 2 inhibitors dexamethasone and hydrocortisone had no major effect. High thermal stability of Od-GST was demonstrated with increasing activity between 4 and 50 °C. Differences between Od-GST and GST of other organisms indicate structural and possibly functional peculiarities and highlight the potential of such enzymes as targets of intervention.
Helene Kringel - One of the best experts on this subject based on the ideXlab platform.
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comparison of innate and th1 type host immune responses in Oesophagostomum dentatum and trichuris suis infections in pigs
Parasite Immunology, 2016Co-Authors: Annette Andreasen, Stig Milan Thamsborg, Elsenoor J Klaver, Helena Mejer, Kerstin Skovgaard, Helene KringelAbstract:Summary This study investigated the details of the innate and Th1/Treg-type-associated host immune responses in Trichuris suis and Oesophagostomum dentatum mono- and co-infected pigs and in vitro in stimulated porcine dendritic cell cultures. Forty-eight pigs were allocated into a 2-factorial design with two groups trickle-inoculated with 10 T. suis eggs/kg/day (Group T) or 20 O. dentatum L3/kg/day (O). Another group (OT) was infected with both parasites. Group C remained uninfected. Expression of innate and Th1/Treg-cell-associated genes in gut mucosa and associated lymph nodes was determined by qPCR at necropsy day 35 and 71. Gene expression showed suppressed/inhibited Th1 and Treg-type immune reactions, in accordance with previous findings of a predominant Th2-type immune response to both nematodes. The in vitro part examined the production of TNF-α in porcine dendritic cells (DC) exposed to T. suis and/or O. dentatum excretory/secretory (E/S) products. Further, binding capacity and structure of E/S products were characterized. Glycan and lectin-binding capacity were generally lower in O. dentatum E/S products compared to T. suis which may explain the earlier found weaker Th2 response to the former. Surprisingly, O. dentatum E/S products induced a significant (P < 0·0001) increase in TNF-α DC production, potentially indicating a new mode of helminth–host immune response interaction.
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Comparison of innate and Th1-type host immune responses in Oesophagostomum dentatum and Trichuris suis infections in pigs
Parasite Immunology, 2015Co-Authors: Annette Andreasen, Stig Milan Thamsborg, Elsenoor J Klaver, Helena Mejer, Kerstin Skovgaard, Helene KringelAbstract:Summary This study investigated the details of the innate and Th1/Treg-type-associated host immune responses in Trichuris suis and Oesophagostomum dentatum mono- and co-infected pigs and in vitro in stimulated porcine dendritic cell cultures. Forty-eight pigs were allocated into a 2-factorial design with two groups trickle-inoculated with 10 T. suis eggs/kg/day (Group T) or 20 O. dentatum L3/kg/day (O). Another group (OT) was infected with both parasites. Group C remained uninfected. Expression of innate and Th1/Treg-cell-associated genes in gut mucosa and associated lymph nodes was determined by qPCR at necropsy day 35 and 71. Gene expression showed suppressed/inhibited Th1 and Treg-type immune reactions, in accordance with previous findings of a predominant Th2-type immune response to both nematodes. The in vitro part examined the production of TNF-α in porcine dendritic cells (DC) exposed to T. suis and/or O. dentatum excretory/secretory (E/S) products. Further, binding capacity and structure of E/S products were characterized. Glycan and lectin-binding capacity were generally lower in O. dentatum E/S products compared to T. suis which may explain the earlier found weaker Th2 response to the former. Surprisingly, O. dentatum E/S products induced a significant (P
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immune and inflammatory responses in pigs infected with trichuris suis and Oesophagostomum dentatum
Veterinary Parasitology, 2015Co-Authors: Annette Andreasen, Helene Kringel, Heidi Huus Petersen, Harry D Dawson, Kerstin Skovgaard, Tine Iburg, Joseph F. Urban, Stig Milan ThamsborgAbstract:The aim of the present study was to investigate parasite induced immune responses in pigs co-infected with Trichuris suis and Oesophagostomum dentatum as compared to mono-species infected pigs. T. suis is known to elicit a strong immune response leading to rapid expulsion, and a strong antagonistic effect on O. dentatum populations has been observed in co-infected pigs. Forty-eight helminth naive pigs were allocated into 4 groups in a 2-factorial design. Two groups were trickle inoculated with either 10 T. suis eggs/kg/day (Group T) or 20 O. dentatum L3/kg/day (Group O). Group OT was infected with same levels of both T. suis and O. dentatum (Group OT) and Group C remained uninfected. In each group, six pigs were necropsied after 35 days and the remaining pigs after 71 days. Parasite E/S-antigen specific serum antibodies were quantified by an in-direct ELISA. qPCR was used to measure the expression of immune function related genes in the mucosa of proximal colon and the draining lymph node. Highly significant interactions were identified for O. dentatum specific IgG1 (p < 0.0001) and IgG2 (p < 0.0006) antibodies with a remarkable 2-fold higher antibody response in group OT pigs as compared to group O. These findings indicated that T. suis enhanced the antibody response against O. dentatum in Group OT. The gene expression data confirmed a strong Type 2 response to T. suis (e.g. marked increase in IL-13, ARG1 and CCL11) and clearly weaker in amplitude and/or delayed onset response to O. dentatum in the single infected group. Interactions were found between the two nematodes with regard to several cytokines, e.g. the increase in IL-13 observed in Group T was absent in Group OT (p = 0.06, proximal colon mucosa, 35 and 71 p.i.). Some of these immune response-related interactions may support, or even partially explain, the observed interactions between the two worm populations in co-infected pigs.
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parasite population dynamics in pigs infected with trichuris suis and Oesophagostomum dentatum
Veterinary Parasitology, 2014Co-Authors: Heidi Huus Petersen, Andreas Roepstorff, Helene Kringel, Annette Andreasen, Stig Milan ThamsborgAbstract:Abstract The aim of the present study was to investigate the population dynamics and potential interactions between Trichuris suis and Oesophagostomum dentatum in experimentally co-infected pigs, by quantification of parasite parameters such as egg excretion, worm recovery and worm location. Forty-eight helminth naive pigs were allocated into four groups. Group O was inoculated with 20 O. dentatum L 3 /kg/day and Group T with 10 T. suis eggs/kg/day. Group OT was inoculated with both 20 O. dentatum L 3 /kg/day and 10 T. suis eggs/kg/day, while Group C was kept as an uninfected control group. All inoculations were trickle infections administered twice weekly and were continued until slaughter. Faecal samples were collected from the rectum of all pigs at day 0, and twice weekly from 2 to 9 weeks post first infection (wpi). Six pigs from each group were necropsied 5 wpi and the remaining 6 pigs from each group were necropsied 10 wpi. The faecal egg counts (FEC) and total worm burdens of O. dentatum were dramatically influenced by the presence of T. suis , with significantly lower mean FECs and worm burdens at 5 and 10 wpi compared to single infected pigs. Furthermore, in the presence of T. suis we found that O. dentatum was located more posteriorly in the gut. The changes in the Trichuris population were less prominent, but faecal egg counts, worm counts 5 wpi (57% recovered vs. 39%) and the proportion of infected animals at 10 wpi were higher in Group OT compared to Group T. The location of T. suis was unaffected by the presence of O. dentatum . These results indicate an antagonistic interaction between T. suis and O. dentatum which is dominated by T. suis .
