Tissue-specific transcriptomes of Anisakis simplex (sensu stricto) and Anisakis pegreffii reveal potential molecular mechanisms involved in pathogenicity.

BACKGROUND: Larval stages of the sibling species of parasitic nematodes Anisakis simplex (sensu stricto) (s.s.) (AS) and Anisakis pegreffii (AP) are responsible for a fish-borne zoonosis, known as anisakiasis, that humans aquire via the ingestion of raw or undercooked infected fish or fish-based products. These two species differ in geographical distribution, genetic background and peculiar traits involved in pathogenicity. However, thus far little is known of key molecules potentially involved in host-parasite interactions. Here, high-throughput RNA-Seq and bioinformatics analyses of sequence data were applied to the characterization of the whole sets of transcripts expressed by infective larvae of AS and AP, as well as of their pharyngeal tissues, in a bid to identify transcripts potentially involved in tissue invasion and host-pathogen interplay. RESULTS: Approximately 34,000,000 single-end reads were generated from cDNA libraries for each species. Transcripts identified in AS and AP encoded 19,403 and 10,424 putative peptides, respectively, and were classified based on homology searches, protein motifs, gene ontology and biological pathway mapping. Differential gene expression analysis yielded 226 and 339 transcripts upregulated in the pharyngeal regions of AS and AP, respectively, compared with their corresponding whole-larvae datasets. These included proteolytic enzymes, molecules encoding anesthetics, inhibitors of primary hemostasis and virulence factors, anticoagulants and immunomodulatory peptides. CONCLUSIONS: This work provides the scientific community with a list of key transcripts expressed by AS and AP pharyngeal tissues and corresponding annotation information which represents a ready-to-use resource for future functional studies of biological pathways specifically involved in host-parasite interplay

The Anisakis Transcriptome Provides a Resource for Fundamental and Applied Studies on Allergy-Causing Parasites.

BACKGROUND: Food-borne nematodes of the genus Anisakis are responsible for a wide range of illnesses (= anisakiasis), from self-limiting gastrointestinal forms to severe systemic allergic reactions, which are often misdiagnosed and under-reported. In order to enhance and refine current diagnostic tools for anisakiasis, knowledge of the whole spectrum of parasite molecules transcribed and expressed by this parasite, including those acting as potential allergens, is necessary. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we employ high-throughput (Illumina) sequencing and bioinformatics to characterise the transcriptomes of two Anisakis species, A. simplex and A. pegreffii, and utilize this resource to compile lists of potential allergens from these parasites. A total of ~65,000,000 reads were generated from cDNA libraries for each species, and assembled into ~34,000 transcripts (= Unigenes); ~18,000 peptides were predicted from each cDNA library and classified based on homology searches, protein motifs and gene ontology and biological pathway mapping. Using comparative analyses with sequence data available in public databases, 36 (A. simplex) and 29 (A. pegreffii) putative allergens were identified, including sequences encoding 'novel' Anisakis allergenic proteins (i.e. cyclophilins and ABA-1 domain containing proteins). CONCLUSIONS/SIGNIFICANCE: This study represents a first step towards providing the research community with a curated dataset to use as a molecular resource for future investigations of the biology of Anisakis, including molecules putatively acting as allergens, using functional genomics, proteomics and immunological tools. Ultimately, an improved knowledge of the biological functions of these molecules in the parasite, as well as of their immunogenic properties, will assist the development of comprehensive, reliable and robust diagnostic tools.This work was supported by a ‘Collaborations Across Boundaries’ grant and a seed grant from the Centre of Biodiscovery and Molecular Development of Therapeutics, James Cook University (FJB and CC). ALL is an Australian Research Council (ARC) Future Fellow and his laboratory is supported by grants from the National Health and Medical Research Council of Australia (NHMRC). Research in the CC laboratory is supported by grants from the Isaac Newton Trust/Wellcome Trust/University of Cambridge (grant number PNVM/GAAB) and the Royal Society (grant number PNAG/428)

Experimental hookworm infection and escalating gluten challenges are associated with increased microbial richness in celiac subjects.

The intestinal microbiota plays a critical role in the development of the immune system. Recent investigations have highlighted the potential of helminth therapy for treating a range of inflammatory disorders, including celiac disease (CeD); however, the mechanisms by which helminths modulate the immune response of the human host and ameliorate CeD pathology are unknown. In this study, we investigated the potential role of alterations in the human gut microbiota in helminth-mediated suppression of an inflammatory disease. We assessed the qualitative and quantitative changes in the microbiota of human volunteers with CeD prior to and following infection with human hookworms, and following challenge with escalating doses of dietary gluten. Experimental hookworm infection of the trial subjects resulted in maintenance of the composition of the intestinal flora, even after a moderate gluten challenge. Notably, we observed a significant increase in microbial species richness over the course of the trial, which could represent a potential mechanism by which hookworms can regulate gluten-induced inflammation and maintain intestinal immune homeostasis.This work was supported by grants 1052938 to C.C., 613718 to P.G., and 1037304 and 1020114 to A.L. from the National Health and Medical Research Council of Australia (NHMRC), and from James Cook University (FMHMS 2013 grants round) and the Isaac Newton Trust / Wellcome Trust ISSF / University of Cambridge Joint Research Grants Scheme to C.C.This is the final version of the article. It first appeared from NPG via http://dx.doi.org/10.1038/srep1379

Divergent Effects of T Cell Costimulation and Inflammatory Cytokine Production on Autoimmune Peripheral Neuropathy Provoked by Aire Deficiency

Chronic Inflammatory Demyelinating Polyneuropathy (CIDP) results from autoimmune destruction of the peripheral nervous system (PNS) and is a component of the multi-organ autoimmunity syndrome which results from Autoimmune Regulator (Aire) gene mutations in humans. In parallel, PNS autoimmunity resembling CIDP develops spontaneously in NOD mice with a partial loss of Aire function (NOD.AireGW/+ mice), and is a T cell-mediated disease. In this study, we analyze how key aspects of T cell activation and function modulate disease development in Aire-deficient mice. We show here that genetic ablation of the Th1 cytokine IFNγ completely prevents clinical and electrophysiological evidence of neuropathy in NOD.AireGW/+ mice. IFNγ deficiency is associated with absence of immune infiltration and decreased expression of the T cell chemoattractant IP-10 in sciatic nerves. Thus, IFNγ is absolutely required for the development of autoimmune peripheral neuropathy in NOD.AireGW/+ mice. Because IFNγ secretion is enhanced by B7-CD28 costimulation of T cells, we sought to determine the effects of these costimulatory molecules on neuropathy development. Surprisingly, B7-2 deficiency accelerated neuropathy development in NOD.AireGW/+ mice, and antibody blockade of both B7-1 and B7-2 resulted in fulminant, early-onset neuropathy. Thus, in contrast to IFNγ, B7-2 alone and B7-1/B7-2 in combination function to ameliorate neuropathy development in NOD.AireGW/+ mice. Together, these findings reveal distinct and opposing effects of T cell costimulatory pathways and IFNγ production on the pathogenesis of autoimmune peripheral neuropathy

Changes in duodenal tissue-associated microbiota following hookworm infection and consecutive gluten challenges in humans with coeliac disease

A reduced diversity of the gastrointestinal commensal microbiota is associated with the development of several inflammatory diseases. Recent reports in humans and animal models have demonstrated the beneficial therapeutic effects of infections by parasitic worms (helminths) in some inflammatory disorders, such as inflammatory bowel disease (IBD) and coeliac disease (CeD). Interestingly, these studies have described how helminths may alter the intestinal microbiota, potentially representing a mechanism by which they regulate inflammation. However, for practical reasons, these reports have primarily analysed the faecal microbiota. In the present investigation, we have assessed, for the first time, the changes in the microbiota at the site of infection by a parasitic helminth (hookworm) and gluten-dependent inflammation in humans with CeD using biopsy tissue from the duodenum. Hookworm infection and gluten exposure were associated with an increased abundance of species within the Bacteroides phylum, as well as increases in the richness and diversity of the tissue-resident microbiota within the intestine, results that are consistent with previous reports using other helminth species in humans and animal models. Hence, this may represent a mechanism by which parasitic helminths may restore intestinal immune homeostasis and exert a therapeutic benefit in CeD, and potentially other inflammatory disorders

The Kuroshio Extension : a leading mechanism for the seasonal sea-level variability along the west coast of Japan

Author Posting. © The Author(s), 2009. This is the author's version of the work. It is posted here by permission of Springer for personal use, not for redistribution. The definitive version was published in Ocean Dynamics 60 (2010): 667-672, doi:10.1007/s10236-009-0239-9.Sea level changes coherently along the two coasts of Japan on the seasonal time scale. AVISO satellite altimetry data and OFES (OGCM for the Earth Simulator) results indicate that the variation propagates clockwise from Japan's east coast through the Tsushima Strait into the Japan/East Sea (JES) and then northward along the west coast. In this study, we hypothesize and test numerically that the sea level variability along the west coast of Japan is remotely forced by the Kuroshio Extension (KE) off the east coast. Topographic Rossby waves and boundary Kelvin waves facilitate the connection. Our 3-d POM model when forced by observed wind stress reproduces well the seasonal changes in the vicinity of JES. Two additional experiments were conducted to examine the relative roles of remote forcing and local forcing. The sea level variability inside the JES was dramatically reduced when the Tsushima Strait is blocked in one experiment. The removal of the local forcing, in another experiment, has little effect on the JES variability. Both experiments support our hypothesis that the open-ocean forcing, possibly through the KE variability, is the leading forcing mechanism for sea level change along the west coast of Japan.This work was conducted when Chao Ma was a visiting graduate student at WHOI. His visit has been supported by China Scholarship Council and WHOI Academics Office. This study has been supported by WHOI’s Coastal Ocean Institute, the National Basic Research Program of China 2005CB422303 and 2007CB481804), the International Science and Technology Cooperation Program of China (2006DFB21250), the Natural Science Foundation of China (40706006) , and the Ministry of Education’s 111 Project (B07036). Lin was supported by the Program for New Century Excellent Talents in University (NECT-07-0781)

An asymmetric upwind flow, Yellow Sea Warm Current : 1. New observations in the western Yellow Sea

Author Posting. © American Geophysical Union, 2011. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Journal of Geophysical Research 116 (2011): C04026, doi:10.1029/2010JC006513.The winter water mass along the Yellow Sea Trough (YST), especially on the western side of the trough, is considerably warmer and saltier than the ambient shelf water mass. This observed tongue-shape hydrographic feature implies the existence of a winter along-trough and onshore current, often referred to as the Yellow Sea Warm Current (YSWC). However, the YSWC has not been confirmed by direct current measurements and therefore skepticism remains regarding its existence. Some studies suggest that the presence of the warm water could be due to frontal instability, eddies, or synoptic scale wind bursts. It is noted that in situ observations used in most previous studies were from the central and eastern sides of the YST even though it is known that the warm water core is more pronounced along the western side. Data from the western side have been scarce. Here we present a set of newly available Chinese observations, including some from a coordinated effort involving three Chinese vessels in the western YST during the 2006–2007 winter. The data show unambiguously the existence of the warm current on the western side of YST. Both the current and hydrography observations indicate a dominant barotropic structure of YSWC. The westward deviation of YSWC axis is particularly obvious to the south of 35°N and is clearly associated with an onshore movement of warm water. To the north of 35°N, the YSWC flows along the bathymetry with slightly downslope movement. We conclude that the barotropic current is mainly responsible for the warm water intrusion, while the Ekman and baroclinic currents play an important but secondary role. These observations help fill an observational gap and establish a more complete view of the YSWC.The authors have been supported by China’s National Basic Research Priorities Programmer (2007CB411804 and 2005CB422303), the Ministry of Education’s 111 Project (B07036), the Program for New Century Excellent Talents in University (NECT‐07‐0781), and the China National Science Fundation (40976004, 40921004. and 40930844). J.Y. is supported by the U.S. National Science Foundation and the Woods Hole Oceanographic Institution’s Coastal Ocean Institute

Infection with the sheep gastrointestinal nematode Teladorsagia circumcincta increases luminal pathobionts

Abstract: Background: The multifaceted interactions between gastrointestinal (GI) helminth parasites, host gut microbiota and immune system are emerging as a key area of research within the field of host-parasite relationships. In spite of the plethora of data available on the impact that GI helminths exert on the composition of the gut microflora, whether alterations of microbial profiles are caused by direct parasite-bacteria interactions or, indirectly, by alterations of the GI environment (e.g. mucosal immunity) remains to be determined. Furthermore, no data is thus far available on the downstream roles that qualitative and quantitative changes in gut microbial composition play in the overall pathophysiology of parasite infection and disease. Results: In this study, we investigated the fluctuations in microbiota composition and local immune microenvironment of sheep vaccinated against, and experimentally infected with, the ‘brown stomach worm’ Teladorsagia circumcincta, a parasite of worldwide socio-economic significance. We compared the faecal microbial profiles of vaccinated and subsequently infected sheep with those obtained from groups of unvaccinated/infected and unvaccinated/uninfected animals. We show that alterations of gut microbial composition are associated mainly with parasite infection, and that this involves the expansion of populations of bacteria with known pro-inflammatory properties that may contribute to the immunopathology of helminth disease. Using novel quantitative approaches for the analysis of confocal microscopy-derived images, we also show that gastric tissue infiltration of T cells is driven by parasitic infection rather than anti-helminth vaccination. Conclusions: Teladorsagia circumcincta infection leads to an expansion of potentially pro-inflammatory gut microbial species and abomasal T cells. This data paves the way for future experiments aimed to determine the contribution of the gut flora to the pathophysiology of parasitic disease, with the ultimate aim to design and develop novel treatment/control strategies focused on preventing and/or restricting bacterial-mediated inflammation upon infection by GI helminths. DCvTgHLrMa4pPTvhPWGjhoVideo Abstrac

Tissue-specific transcriptomes of Anisakis simplex (sensu stricto) and Anisakis pegreffii reveal potential molecular mechanisms involved in pathogenicity

BACKGROUND: Larval stages of the sibling species of parasitic nematodes Anisakis simplex (sensu stricto) (s.s.) (AS) and Anisakis pegreffii (AP) are responsible for a fish-borne zoonosis, known as anisakiasis, that humans aquire via the ingestion of raw or undercooked infected fish or fish-based products. These two species differ in geographical distribution, genetic background and peculiar traits involved in pathogenicity. However, thus far little is known of key molecules potentially involved in host-parasite interactions. Here, high-throughput RNA-Seq and bioinformatics analyses of sequence data were applied to the characterization of the whole sets of transcripts expressed by infective larvae of AS and AP, as well as of their pharyngeal tissues, in a bid to identify transcripts potentially involved in tissue invasion and host-pathogen interplay. RESULTS: Approximately 34,000,000 single-end reads were generated from cDNA libraries for each species. Transcripts identified in AS and AP encoded 19,403 and 10,424 putative peptides, respectively, and were classified based on homology searches, protein motifs, gene ontology and biological pathway mapping. Differential gene expression analysis yielded 226 and 339 transcripts upregulated in the pharyngeal regions of AS and AP, respectively, compared with their corresponding whole-larvae datasets. These included proteolytic enzymes, molecules encoding anesthetics, inhibitors of primary hemostasis and virulence factors, anticoagulants and immunomodulatory peptides. CONCLUSIONS: This work provides the scientific community with a list of key transcripts expressed by AS and AP pharyngeal tissues and corresponding annotation information which represents a ready-to-use resource for future functional studies of biological pathways specifically involved in host-parasite interplay

Genome-Wide Characterization of <i>MADS-box</i> Genes Identifies Candidates Associated with Flower and Fruit Development in Loquat (<i>Eriobotrya japonica</i> Lindl.)

The MADS-box transcription factors have garnered substantial attention due to their crucial involvement in various biological processes, particularly in flower organogenesis. A comprehensive investigation into the MADS-box genes remains lacking in loquat (Eriobotrya japonica Lindl.). In the current study, to preliminarily explore the potential candidate genes related to flower and fruit development, a genome-wide analysis was carried out to identify and characterize the MADS-box gene family in loquat. Among the 125 identified EjMADS-box members, 49 genes belonged to type Ⅰ, which were subsequently assigned to three subfamilies: Mα (25 genes), Mβ (10 genes), and Mγ (14 genes). Additionally, 76 genes fell under type II, which were categorized into two groups: MIKCC (70 genes) and MIKC* (6 genes). Through the collinearity analysis and comparison of the gene numbers between loquat and other Rosaceae genomes, it was revealed that the type Ⅱ MADS-box members were expanded in Maloideae after a whole genome duplication. The gene expression analysis utilizing various tissues during flower development revealed that the expression patterns of the ABCDE model homologs were conserved in loquat. In addition, several candidate genes potentially involved in flower bud differentiation (EjMADS107/109) and fruit expansion (EjMADS24/46/49/55/61/67/77/86) were identified. This analysis could serve as a fundamental basis for investigating the molecular functions of the MADS-box genes in the development of flowers as well as fruits in loquat