Extracellular vesicles from parasitic helminths contain specific excretory/secretory proteins and are internalized in intestinal host cells.

The study of host-parasite interactions has increased considerably in the last decades, with many studies focusing on the identification of parasite molecules (i.e. surface or excretory/secretory proteins (ESP)) as potential targets for new specific treatments and/or diagnostic tools. In parallel, in the last few years there have been significant advances in the field of extracellular vesicles research. Among these vesicles, exosomes of endocytic origin, with a characteristic size ranging from 30-100 nm, carry several atypical secreted proteins in different organisms, including parasitic protozoa. Here, we present experimental evidence for the existence of exosome-like vesicles in parasitic…

A bug's life: Delving into the challenges of helminth microbiome studies.

The body of vertebrates is inhabited by trillions of microorganisms, i.e. viruses, archaea, bacteria and unicellular eukaryotes, together referred to as the ‘microbiota’. Similarly, vertebrates also host a plethora of parasitic worms (the ‘macrobiota’), some of which share their environment with the microbiota inhabiting the gastrointestinal tract [1]. Complex interactions between the helminths and the gut microbiota have been associated with establishment of parasite infection, disease manifestations, and host immune-modulation [2, 3]. Remarkably, not only enteric helminths alter the 26 gut microbiome composition [4], but also the infection with blood flukes of the genus Schistosoma has be…

Baseline Gut Microbiota Composition Is Associated With Schistosoma mansoni Infection Burden in Rodent Models

In spite of growing evidence supporting the occurrence of complex interactions between Schistosoma and gut bacteria in mice and humans, no data is yet available on whether worm-mediated changes in microbiota composition are dependent on the baseline gut microbial profile of the vertebrate host. In addition, the impact of such changes on the susceptibility to, and pathophysiology of, schistosomiasis remains largely unexplored. In this study, mice colonized with gut microbial populations from a human donor (HMA mice), as well as microbiota-wild type (WT) animals, were infected with Schistosoma mansoni, and alterations of their gut microbial profiles at 50 days post-infection were compared to …

Definitive host influences the proteomic profile of excretory/secretory products of the trematode Echinostoma caproni

Background Echinostoma caproni is an intestinal trematode extensively used as experimental model for the study of factors that determine the course of intestinal helminth infections, since this markedly depends on the host species. Although the host-dependent mechanisms for either chronic establishment or early parasite rejection have been broadly studied, little is known regarding the parasite response against different host environments. Methods To identify host-dependent differentially expressed proteins, a comparative proteomic analysis of the excretory/secretory products released from E. caproni adults, isolated from hosts displaying different compatibility with this trematode, was per…

Th2 and Th1 Responses: Clear and Hidden Sides of Immunity Against Intestinal Helminths.

Intestinal helminthiases affect millions of people worldwide, mainly in developing regions, where they cause a significant negative impact on human health and socioeconomic growth of affected populations. However, intestinal helminthiases are still among the most neglected tropical diseases. Protective immunity against intestinal helminths is associated with development of type 2 responses. Nevertheless, in some host–intestinal helminth combinations, local Th1 responses are initiated, inducing chronicity. The usage of helminth–mouse models is useful for elucidating the mechanisms behind the initiation of each type of response. Herein, the current knowledge on these topics is reviewed, payin…

Additional file 1: of Definitive host influences the proteomic profile of excretory/secretory products of the trematode Echinostoma caproni

Detailed gel images of the differential spots identified by mass spectrometry and database search. Magnification of the 2D gel-areas corresponding to the 19 differentially expressed spots identified by mass spectrometry and database search. Detailed images for each of the 6 replicates analyzed (3 corresponding to the ESPs of E. caproni adults obtained from rats, and 3 from those isolated from mice) are shown. Spot numbers refer to gel image in Fig. 2. (PPTX 4834 kb)

Additional file 2: of Definitive host influences the proteomic profile of excretory/secretory products of the trematode Echinostoma caproni

Details of the computational comparison of the excretory/secretory proteomes. Description of data: Quantitative and statistical details of the computational comparison of excretory/secretory proteomes of Echinostoma caproni adult worms isolated from mice and rats using Progenesis SameSpots software (version 4.5) (Nonlinear Dynamics Ltd.). Spot numbers refer to gel image in Fig. 2. (DOCX 13 kb)

Differential alterations in the small intestine epithelial cell turnover during acute and chronic infection with Echinostoma caproni (Trematoda)

Background The intestinal epithelium plays a multifactorial role in mucosal defense. In this sense, augmented epithelial cell turnover appears as a potential effector mechanism for the rejection of intestinal-dwelling helminths. Methods A BrdU pulse-chase experiment was conducted to investigate the infection-induced alterations on epithelial cell kinetics in hosts of high (mouse) and low (rat) compatibility with the intestinal trematode Echinostoma caproni. Results High levels of crypt-cell proliferation and tissue hyperplasia were observed in the ileum of infected mice, coinciding with the establishment of chronic infections. In contrast, the cell migration rate was about two times higher …

Proteomic analysis of the pinworm Syphacia muris (Nematoda: Oxyuridae), a parasite of laboratory rats

Syphacia muris (Nematoda: Oxyuridae) is a ubiquitous nematode that commonly infects rats in the laboratory which can interfere in the development of biological assays. The somatic extract of S. muris adults collected from infected rats was investigated using a proteomic approach. A shot-gun liquid chromatography/tandem mass spectrometry procedure was used. We used the MASCOT search engine (Matrix-Science) and ProteinPilot software v2.0 (Applied Biosystems) for the database search. A total of 359 proteins were accurately identified from the worms. The largest protein families consisted of metabolic enzymes and those involved in the nucleic metabolism and cell cycle. Proteins of transmembrane…

New Insights into the Treatment of Foodborne Trematode Infections

Helminths, hosts, and their microbiota: new avenues for managing gastrointestinal helminthiases in ruminants

Evidence is emerging of complex interactions occurring between gastrointestinal (GI) parasites of ruminants and the resident gut flora, with likely implications for the pathophysiology of worm infection and disease. Similarly, recent data point toward the occurrence of a GI nematode (GIN)-specific microbiota, with potential roles in worm fundamental physiology and reproduction. Parasite-microbiota relationships might represent potential targets for the development of novel parasiticides.In this article, we review current knowledge of the role(s) that host- and helminth-associated microbiota play in ruminant host-parasite relationships, and outline potential avenues for the control of GIN of…

Additional file 4: of Definitive host influences the proteomic profile of excretory/secretory products of the trematode Echinostoma caproni

Biological process-based classification of proteins overexpressed in mice. Significantly overexpressed proteins in the excretory/secretory products of Echinostoma caproni adults obtained from mice, classified according to their Gene Ontology (GO)-predicted biological process. Pie chart represents the number of proteins assigned to each GO category (biological process, level 4). Proteins included in each category are listed below the graph. (PPTX 118 kb)

Helminth Microbiota Profiling Using Bacterial 16S rRNA Gene Amplicon Sequencing: From Sampling to Sequence Data Mining

Symbiont microbial communities play important roles in animal biology and are thus considered integral components of metazoan organisms, including parasitic worms (helminths). Nevertheless, the study of helminth microbiomes has thus far been largely overlooked, and symbiotic relationships between helminths and their microbiomes have been only investigated in selected parasitic worms. Over the past decade, advances in next-generation sequencing technologies, coupled with their increased affordability, have spurred investigations of helminth-associated microbial communities aiming at enhancing current understanding of their fundamental biology and physiology, as well as of host-microbe intera…

Resistance against Echinostoma caproni (Trematoda) secondary infections in mice is not dependent on the ileal protein production

Echinostoma caproni (Trematoda: Echinostomatidae) is an intestinal trematode, which has been widely employed to investigate the factors determining the rejection of intestinal helminths. Protein production patterns of intestinal epithelial cells are related to the infection-induced changes that determine the course of E. caproni infections. Herein, we compare the protein production profiles in the ileum of four experimental groups of mice: control; infected; dewormed and reinfected. Worm burdens were significantly lower in secondary infections, confirming the generation of partial resistance to homologous secondary infections in mice. However, quantitative comparison by 2D-DIGE showed that …

Th17 responses in Echinostoma caproni infections in hosts of high and low compatibility.

In order to investigate the factors determining the expulsion of intestinal helminths, we have analyzed the in vivo expression of IL-17, TGF-β and IL-23 in several tissues of two host species displaying different compatibility with Echinostoma caproni (Trematoda). We did not observe upregulation of these cytokines in any of the tissues of the high compatible host (mice). In contrast, the responses in the host of low compatibility (rats) with the parasite were markedly different. Significant increases in the expression of IL-17 and TGF-β were observed in the Peyer's patches and the intestine from the 2 to 8 weeks post-infection. The expression of IL-23 was upregulated from 2 to 4 weeks post-…

Protective immunity against Echinostoma caproni in rats is induced by Syphacia muris infection.

Syphacia muris (Nematoda: Oxyuridae) is a ubiquitous nematode that commonly infects rats in the laboratory and can interfere with the development of unrelated biological assays. In this context, we analysed the effect of a patent S. muris infection in Wistar rats on a superimposed infection with the intestinal trematode, Echinostoma caproni (Trematoda: Echinostomatidae). The results indicate that in the rats, infection with S. muris induces an immunity against a subsequent infection with E. caproni, although each parasite occupies different niches in the host. Echinostoma caproni worm recovery was significantly decreased in the rats primarily infected with S. muris and, at 3 and 4 weeks pos…

The effect of glycosylation of antigens on the antibody responses against Echinostoma caproni (Trematoda: Echinostomatidae).

SUMMARYIn the present study, we analyse the effect of glycosylation inEchinostoma caproni(Trematoda: Echinostomatidae) antigens in antibody responses against the parasite in experimentally infected mice. It has been previously demonstrated that the mouse is a host of high compatibility withE. caproniand develops elevated responses of IgG, IgG1, IgG3 and IgM as a consequence of the infection, though the role of glycans in these responses remains unknown. To this purpose, the responses generated in mice against non-treated excretory/secretory antigens ofE. caproniwere compared with those observed after N-deglycosylation, O-deglycosylation and double deglycosylation of the antigens by indirect…

Echinostoma caproni (Trematoda): differential in vivo mucin expression and glycosylation in high- and low-compatible hosts.

Enhanced mucus production and release appears to be a common mechanism for the clearance of intestinal helminths, and this expulsion is normally mediated by Th2-type immune responses. To investigate the factors determining the expulsion of intestinal helminths, we have analysed in vivo expression of mucin genes at the site of infection in two host species displaying different compatibility with Echinostoma caproni (Trematoda). Surprisingly, a general down-regulation on mucin mRNA expression was detected in low-compatible hosts (rats) coinciding with the development of Th2/Th17 responses and the early rejection of the worms from the intestinal lumen. This suggests the existence of a mechanis…

Altered Protein Expression in the Ileum of Mice Associated with the Development of Chronic Infections with Echinostoma caproni (Trematoda)

Background Echinostoma caproni (Trematoda: Echinostomatidae) is an intestinal trematode that has been extensively used as experimental model to investigate the factors determining the expulsion of intestinal helminths or, in contrast, the development of chronic infections. Herein, we analyze the changes in protein expression induced by E. caproni infection in ICR mice, a host of high compatibility in which the parasites develop chronic infections. Methodology/Principal Findings To determine the changes in protein expression, a two-dimensional DIGE approach using protein extracts from the intestine of naïve and infected mice was employed; and spots showing significant differential expression…

Additional file 5: of Definitive host influences the proteomic profile of excretory/secretory products of the trematode Echinostoma caproni

Molecular function-based classification of proteins overexpressed in mice. Significantly overexpressed proteins in the excretory/secretory products of Echinostoma caproni adults obtained from mice, classified according to their Gene Ontology (GO)-predicted molecular function. Pie chart represents the number of proteins assigned to each GO category (molecular function, level 3). Proteins included in each category are listed below the graph. (PPTX 77 kb)

Classic Models for New Perspectives: Delving into Helminth–Microbiota–Immune System Interactions

Whilst a wealth of data indicate that infections by gastrointestinal helminths are accompanied by significant alterations in the composition of the vertebrate gut flora, little is known of the immune-molecular mechanisms that regulate host-parasite-microbiota interactions. 'Traditional' experimental models of gastrointestinal helminthiases, in which the role(s) of each of the components of this triad can be tested, provide an opportunity to advance research in this area. In this article, we propose the Echinostoma caproni-mouse system as a potentially useful tool for studies of the role of the host gut microbiota in preventing pathology and inducing parasite clearance via interleukin (IL)-2…

Partial resistance to homologous challenge infections of the digenean Echinostoma caproni in ICR mice

AbstractIn the present paper, we analyse the effect of a primary infection of ICR mice with Echinostoma caproni (Trematoda: Echinostomatidae) on the generation of resistance against homologous challenge infections. In ICR mice, E. caproni induces chronic infections concomitantly with strong responses characterized by the development of T-helper 1 (Th1)-type local immune responses with elevated levels of local interferon-gamma (IFN-γ) and inflammatory and antibody responses. Here, the effect of the response generated against a primary infection with E. caproni in the generation of resistance against subsequent homologous infections was analysed. For this purpose, ICR mice were challenged wit…

Additional file 3: of Definitive host influences the proteomic profile of excretory/secretory products of the trematode Echinostoma caproni

Identification details of differentially expressed proteins. Description of data: Details of the identification of differentially expressed proteins using X!Tandem and MS-GF+ search engines on the Echinostoma caproni genome and transcriptome databases and BLASTp analysis against NCBInr protein database. Spot numbers refer to gel image in Fig. 2. (DOCX 19 kb)

Parásitos intestinales. Helmintiasis

Differential expression and glycosylation of proteins in the rat ileal epithelium in response to Echinostoma caproni infection.

Echinostoma caproni (Trematoda: Echinostomatidae) is an intestinal trematode that has been used as experimental model to investigate the factors determining the expulsion of intestinal helminths. We analyze the changes in the protein expression and glycosylation induced by E. caproni in Wistar rat, a host of low compatibility in which the parasites are rapidly rejected. To determine the changes in protein expression, two-dimensional difference gel electrophoresis was employed using protein extracts from the intestine of naïve and infected rats. The patterns of glycosylation were analyzed by lectin blotting. Those spots showing differential expression or glycosylation were analyzed by mass s…

Antibody trapping: A novel mechanism of parasite immune evasion by the trematode Echinostoma caproni

Background Helminth infections are among the most prevalent neglected tropical diseases, causing an enormous impact in global health and the socioeconomic growth of developing countries. In this context, the study of helminth biology, with emphasis on host-parasite interactions, appears as a promising approach for developing new tools to prevent and control these infections. Methods/Principal findings The role that antibody responses have on helminth infections is still not well understood. To go in depth into this issue, work on the intestinal helminth Echinostoma caproni (Trematoda: Echinostomatidae) has been undertaken. Adult parasites were recovered from infected mice and cultured in vi…

Interleukin-25 Induces Resistance Against Intestinal Trematodes

AbstractEchinostoma caproni is an intestinal trematode that has been extensively used as an experimental model to investigate the factors determining the resistance to intestinal helminths or the development of chronic infections. ICR mice are permissive hosts for E. caproni in which chronic infections are developed, concomitantly with local Th1 responses, elevated levels of local IFN-γ, inflammation and antibody responses. However, mice develop partial resistance to homologous challenge infections after cure of a primary infection, which converts this subject into an adequate model for the study of the mechanisms generating resistance against intestinal helminths. The purpose of the presen…

Gut-microbiota-derived extracellular vesicles: Overlooked mediators in host–helminth interactions?

Helminth infections impact the composition of the mammalian gut microbiota; however, the mechanisms underpinning these interactions are, thus far, unknown. In this article, we propose that microbiota-derived extracellular vesicles might represent key players in host-helminth-microbiome crosstalk, and outline future directions to elucidate their role(s) in host-parasite relationships.