0000000001110416
AUTHOR
Javier Sotillo
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…
Echinostoma caproni: Differential tegumental responses to growth in compatible and less compatible hosts
Abstract The topography of the tegument of Echinostoma caproni adults collected from high (mice) and low (rats) compatible hosts was compared by SEM. In the oral (OS) and the ventral sucker (VS) areas, a worm age–host species interaction was found with regard to the density of spines. There was a decrease in the density of spines in the adults collected from mice, whereas an increase occurred in the OS area in worms from rats over time. The tegumentary spines in adults from mice became larger and blunter. Some spines from the VS area in adults from mice at 4 wpi were multipointed. The spines of adults from rats were sharper, not covered by the tegument and no multipointed spines were observ…
The protein and microRNA cargo of extracellular vesicles from parasitic helminths – current status and research priorities
Helminth parasites have a remarkable ability to persist within their mammalian hosts, which is largely due to their secretion of molecules with immunomodulatory properties. Although the soluble components of helminth secretions have been extensively studied, the discovery that helminths release extracellular vesicles (EVs) has added further complexity to the host-parasite interaction. Whilst several studies have begun to characterise the molecules carried by helminth EVs, work aimed at investigating their biological functions has been hindered by a lack of helminth-specific EV markers. To begin to address this, we summarised helminth EV literature to date. With a focus on the protein and mi…
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…
A QUANTITATIVE APPROACH TO THE EXPERIMENTAL TRANSMISSION SUCCESS OF ECHINOSTOMA FRIEDI (TREMATODA: ECHINOSTOMATIDAE) IN RATS
Using a range of parameters, the ability of rats (Rattus norvegicus) to successfully transmit Echinostoma friedi to the next host was examined under experimental conditions. The concept of Experimental Transmission Success (TM), defined as the number of hosts that become successfully infected after exposure to a number of infective stages produced by a previous host per unit of inoculation at which this latter host was exposed, was introduced. Using data for the egg output and miracidium hatching and infectivity, the TM permits us to estimate the ability of a particular defintive host species to successfully transmit a parasite species. This concept may be also useful to compare the transmi…
Exploiting Helminth–Host Interactomes through Big Data
Helminths facilitate their parasitic existence through the production and secretion of different molecules, including proteins. Some helminth proteins can manipulate the host's immune system, a phenomenon that is now being exploited with a view to developing therapeutics for inflammatory diseases. In recent years, hundreds of helminth genomes have been sequenced, but as a community we are still taking baby steps when it comes to identifying proteins that govern host-helminth interactions. The information generated from genomic, immunomic, and proteomic studies, as well as from cutting-edge approaches such as proteogenomics, is leading to a substantial volume of big data that can be utilised…
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)
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…
Minimal information for studies of extracellular vesicles 2018 (MISEV2018):a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines
The last decade has seen a sharp increase in the number of scientific publications describing physiological and pathological functions of extracellular vesicles (EVs), a collective term covering various subtypes of cell-released, membranous structures, called exosomes, microvesicles, microparticles, ectosomes, oncosomes, apoptotic bodies, and many other names. However, specific issues arise when working with these entities, whose size and amount often make them difficult to obtain as relatively pure preparations, and to characterize properly. The International Society for Extracellular Vesicles (ISEV) proposed Minimal Information for Studies of Extracellular Vesicles ("MISEV") guidelines fo…
Development and pathology of echinostoma caproni in experimentally infected mice
In the present article, several parasitological features of mice, each experimentally infected with 75 metacercariae of Echinostoma caproni (Trematoda: Echinostomatidae), were studied during the first 12 wk postinfection. Moreover, the early pathological responses also were analyzed and compared with data previously published on other host species of E. caproni to gain further insight into the factors determining worm rejection or establishment of chronic infections. The results obtained show that the pattern of E. caproni infection in mice is consistent with a highly compatible host–parasite system. This combination is characterized by a high worm establishment, high egg output, and long s…
Echinostoma caproni (Trematoda): differential in vivo cytokine responses in high and low compatible hosts.
In order to investigate the factors determining the expulsion of intestinal trematodes, we have analyzed the in vivo cytokine responses at several levels and the local responses against Echinostoma caproni (Trematoda) in two host species displaying different compatibility with the parasite. The response of the high compatible host (mice) is characterized by a mixed Th1/Th2 phenotype in the spleen, Peyer's patches and mesenteric lymph nodes. At the intestine, a marked Th1 response with a marked increase of IFN-γ together with elevated number of mucosal neutrophils and expression of induced nitric oxide synthase were observed. The responses in the host of low compatibility (rats) with the par…
Analysis of the tegument of Zygocotyle lunata (Trematoda: Paramphistomidae) adults by scanning electron microscopy.
Paramphistomiasis is a parasitic disease caused by various species of the Paramphistomidae. These species mainly affect domestic and wild ruminants; the economic impact of these diseases is often underestimated. Traditionally, the identification of paramphistomes has been difficult and has been based on morphological aspects such as the body shape, the position of the esophagus and cecae, or the tegumental papillae. Despite the many investigations regarding the tegumental papillae for other paramphistomes, very few efforts have been made using Zygocotyle lunata, partly because many authors differ with respect to the importance of papillae in the classification of paramphistomids. Herein, we…
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)
Echinostoma caproni: kinetics of IgM, IgA and IgG subclasses in the serum and intestine of experimentally infected rats and mice.
The kinetics of specific immunoglobulin M, A and IgG subclasses against Echinostoma caproni (Trematoda: Echinostomatidae) were analyzed in serum and intestinal fluid of two host species (Wistar rats and ICR mice) in which the course of the infection markedly differs. In rats, the worms were rapidly expelled, whereas E. caproni evokes in mice long-lasting infection. The pattern of antibody responses in both serum and intestinal samples was different in each host species. Serum responses in mice were characterized by significant increases of IgM, IgA, total IgG, IgG1 and IgG3, but not IgG2a. In contrast, serum responses in rats showed elevated levels of IgM, probably in relation to thymus-ind…
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 …
The transcriptome analysis of Strongyloides stercoralis L3i larvae reveals targets for intervention in a neglected disease.
Background: Strongyloidiasis is one of the most neglected diseases distributed worldwide with endemic areas in developed countries, where chronic infections are life threatening. Despite its impact, very little is known about the molecular biology of the parasite involved and its interplay with its hosts. Next generation sequencing technologies now provide unique opportunities to rapidly address these questions. Principal Findings: Here we present the first transcriptome of the third larval stage of S. stercoralis using 454 sequencing coupled with semi-automated bioinformatic analyses. 253,266 raw sequence reads were assembled into 11,250 contiguous sequences, most of which were novel. 8037…
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-…
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…
Zygocotyle lunata: Proteomic analysis of the adult stage
The somatic extract of Zygocotyle lunata (Trematoda: Paramphistomidae) adults collected from experimentally infected mice was investigated using a proteomic approach to separate and identify tryptic peptides from the somatic extract of Z. lunata adult worms. 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 36 proteins were accurately identified from the worms. The largest protein family consisted of metabolic enzymes. Structural, motor and receptor binding proteins and proteins related to oxygen transport were identified in th…
Proteomic analysis of Strongyloides stercoralis L3 larvae
SUMMARYStrongyloidiasis can be perpetuated by autoinfection with the filariform larvae L3, causing asymptomatic chronic infections and creating a population of carriers, affecting not only developing countries. So far, very little is known about the proteins that interact with the human host, and few proteins from the infective Strongyloides stercoralis L3 have been characterized. Here, we report results obtained from a proteomic analysis of the proteins from S. stercoralis L3 larvae obtained from patients. Since the genome of S. stercoralis is not yet available, we used proteomic analysis to identify 26 different proteins, 13 of them released by short digestion with trypsin, which could re…
Leucine aminopeptidase is an immunodominant antigen of Fasciola hepatica excretory and secretory products in human infections.
ABSTRACT The liver fluke Fasciola hepatica parasitizes humans and ruminant livestock worldwide, and it is now being considered a reemerging zoonotic disease, especially in areas in which it is endemic, such as South America. This study investigates the immune response to excretory and secretory products produced by F. hepatica in a group of patients from the Peruvian Altiplano, where the disease is highly endemic. Using a proteomic approach and immunoblotting techniques, we have identified the enzymes leucine aminopeptidase (LAP) and phosphoenolpyruvate carboxykinase as immunodominant antigens recognized by sera from fasciolosis patients. An indirect enzyme-linked immunosorbent assay using …
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…
Special considerations for studies of extracellular vesicles from parasitic helminths: A community-led roadmap to increase rigour and reproducibility.
Over the last decade, research interest in defining how extracellular vesicles (EVs) shape cross-species communication has grown rapidly. Parasitic helminths, worm species found in the phyla Nematoda and Platyhelminthes, are well-recognised manipulators of host immune function and physiology. Emerging evidence supports a role for helminth-derived EVs in these processes and highlights EVs as an important participant in cross-phylum communication. While the mammalian EV field is guided by a community-agreed framework for studying EVs derived from model organisms or cell systems [e.g., Minimal Information for Studies of Extracellular Vesicles (MISEV)], the helminth community requires a supplem…
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)
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)
Identification of antigenic proteins from Echinostoma caproni (Trematoda) recognized by mouse immunoglobulins M, A and G using an immunoproteomic approach.
Antigenic proteins of Echinostoma caproni (Trematoda) against mouse IgM, IgA, IgG, IgG1 and IgG2a were investigated by immunoproteomics. Excretory/secretory products (ESP) of E. caproni separated by two-dimensional (2D) gel electrophoresis were transferred to nitrocellulose membranes and probed with the different mouse immunoglobulin classes. A total of four proteins (enolase, 70 kDa heat-shock protein (HSP-70), actin and aldolase) were accurately identified. Enolase was recognized in eight different spots of which seven of them were detected in the expected molecular weight and were recognized by IgA, IgG or IgG and IgG1. Another spot identified as enolase at 72 kDa was only recognized by …
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.