scholarly journals Recognition Pattern of the Fasciola hepatica Excretome/Secretome during the Course of an Experimental Infection in Sheep by 2D Immunoproteomics

Pathogens ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 725
Author(s):  
David Becerro-Recio ◽  
Javier González-Miguel ◽  
Alberto Ucero ◽  
Javier Sotillo ◽  
Álvaro Martínez-Moreno ◽  
...  
Keyword(s):  
Experimental Infection ◽  
Fasciola Hepatica ◽  
Cathepsin L ◽  
Helminth Parasites ◽  
Glutathione S Transferase ◽  
Serum Samples ◽  
Secretory Products ◽  
Immunogenic Proteins ◽  
Host Parasite ◽  
First Time

Excretory/secretory products released by helminth parasites have been widely studied for their diagnostic utility, immunomodulatory properties, as well as for their use as vaccines. Due to their location at the host/parasite interface, the characterization of parasite secretions is important to unravel the molecular interactions governing the relationships between helminth parasites and their hosts. In this study, the excretory/secretory products from adult worms of the trematode Fasciola hepatica (FhES) were employed in a combination of two-dimensional electrophoresis, immunoblot and mass spectrometry, to analyze the immune response elicited in sheep during the course of an experimental infection. Ten different immunogenic proteins from FhES recognized by serum samples from infected sheep at 4, 8, and/or 12 weeks post-infection were identified. Among these, different isoforms of cathepsin L and B, peroxiredoxin, calmodulin, or glutathione S-transferase were recognized from the beginning to the end of the experimental infection, suggesting their potential role as immunomodulatory antigens. Furthermore, four FhES proteins (C2H2-type domain-containing protein, ferritin, superoxide dismutase, and globin-3) were identified for the first time as non-immunogenic proteins. These results may help to further understand host/parasite relationships in fasciolosis, and to identify potential diagnostic molecules and drug target candidates of F. hepatica.

scholarly journals Identification of Myoferlin, a Potential Serodiagnostic Antigen of Clonorchiasis, via Immunoproteomic Analysis of Sera From Different Infection Periods and Excretory-Secretory Products of Clonorchis sinensis

2021 ◽  
Vol 11 ◽  
Author(s):  
Xiao-Xiao Ma ◽  
Yang-Yuan Qiu ◽  
Zhi-Guang Chang ◽  
Jun-Feng Gao ◽  
Rui-Ruo Jiang ◽  
...  
Keyword(s):  
Light Chain ◽  
Fasciola Hepatica ◽  
Clonorchis Sinensis ◽  
Dynein Light Chain ◽  
Liquid Chromatography Tandem Mass ◽  
Causative Agents ◽  
Specific Proteins ◽  
Secretory Products ◽  
Negative Controls ◽  
Host Parasite

Clonorchiasis, which is caused by Clonorchis sinensis, is an important foodborne disease worldwide. The excretory-secretory products (ESPs) of C. sinensis play important roles in host-parasite interactions by acting as causative agents. In the present study, the ESPs and sera positive for C. sinensis were collected to identify proteins specific to the sera of C. sinensis (i.e., proteins that do not cross-react with Fasciola hepatica and Schistosoma japonicum) at different infection periods. Briefly, white Japanese rabbits were artificially infected with C. sinensis, and their sera were collected at 7 days post-infection (dpi), 14 dpi, 35 dpi, and 77 dpi. To identify the specific proteins in C. sinensis, a co-immunoprecipitation (Co-IP) assay was conducted using shotgun liquid chromatography tandem-mass spectrometry (LC-MS/MS) to pull down the sera roots of C. sinensis, F. hepatica, and S. japonicum. For the annotated proteins, 32, 18, 39, and 35 proteins specific to C. sinensis were pulled down by the infected sera at 7, 14, 35, and 77 dpi, respectively. Three proteins, Dynein light chain-1, Dynein light chain-2 and Myoferlin were detected in all infection periods. Of these proteins, myoferlin is known to be overexpressed in several human cancers and could be a promising biomarker and therapeutic target for cancer cases. Accordingly, this protein was selected for further studies. To achieve a better expression, myoferlin was truncated into two parts, Myof1 and Myof2 (1,500 bp and 810 bp), based on the antigenic epitopes provided by bioinformatics. The estimated molecular weight of the recombinant proteins was 57.3 ku (Myof1) and 31.3 ku (Myof2). Further, both Myof1 and Myof2 could be probed by the sera from rabbits infected with C. sinensis. No cross-reaction occurred with the positive sera of S. japonica, F. hepatica, and negative controls. Such findings indicate that myoferlin may be an important diagnostic antigen present in the ESPs. Overall, the present study provides new insights into proteomic changes between ESPs and hosts in different infection periods by LC-MS/MS. Moreover, myoferlin, as a biomarker, may be used to develop an objective method for future diagnosis of clonorchiasis.

scholarly journals Characterisation of Trichuris muris secreted proteins and extracellular vesicles provides new insights into host-parasite communication

2017 ◽  
Author(s):  
Ramon M. Eichenberger ◽  
Md Hasanuzzaman Talukder ◽  
Matthew A. Field ◽  
Phurpa Wangchuk ◽  
Paul Giacomin ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Control Measures ◽  
Host Cells ◽  
Parasitic Nematodes ◽  
Trichuris Muris ◽  
Future Studies ◽  
Secretory Products ◽  
Host Parasite ◽  
First Time ◽  
Host Tissues

AbstractWhipworms are parasitic nematodes that live in the gut of more than 500 million people worldwide. Due to the difficulty in obtaining parasite material, the mouse whipworm Trichuris muris has been extensively used as a model to study human whipworm infections. These nematodes secrete a multitude of compounds that interact with host tissues where they orchestrate a parasitic existence. Herein we provide the first comprehensive characterisation of the excretory/secretory products of T. muris. We identify 148 proteins secreted by T. muris and show for the first time that the mouse whipworm secretes exosome-like extracellular vesicles (EVs) that can interact with host cells. We use an Optiprep® gradient to purify the EVs, highlighting the suitability of this method for purifying EVs secreted by a parasitic nematode. We also characterise the proteomic and genomic content of the EVs, identifying >350 proteins, 56 miRNAs (22 novel) and 475 full-length mRNA transcripts mapping to T. muris gene models. Many of the miRNAs putatively mapped to mouse genes involved in regulation of inflammation, implying a role in parasite-driven immunomodulation. In addition, for the first time to our knowledge, we use colonic organoids to demonstrate the internalisation of parasite EVs by host cells. Understanding how parasites interact with their host is crucial to develop new control measures. This first characterisation of the proteins and EVs secreted by T. muris provides important information on whipworm-host communication and forms the basis for future studies.

scholarly journals SARS-CoV-2 Seroprevalence in Household Domestic Ferrets (Mustela putorius furo)

Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 667
Author(s):  
Jacobo Giner ◽  
Sergio Villanueva-Saz ◽  
Ana Pilar Tobajas ◽  
María Dolores Pérez ◽  
Ana González ◽  
...  
Keyword(s):  
Experimental Infection ◽  
Animal Species ◽  
Enzyme Linked Immunosorbent Assay ◽  
Receptor Binding Domain ◽  
Mustela Putorius ◽  
Natural Conditions ◽  
Serum Samples ◽  
Source Of Infection ◽  
First Time ◽  
Mustela Putorius Furo

Animal infections with SARS-CoV-2 have been reported in different countries and several animal species have been proven to be susceptible to infection with SARS-CoV-2 both naturally and by experimental infection. Moreover, infections under natural conditions in more than 20 mink farms have been reported where humans could have been the source of infection for minks. However, little information is available about the susceptibility of pet animals under natural conditions and currently there is no SARS-CoV-2 epidemiological assessment occurrence in household ferrets. In this study, the presence of SARS-CoV-2 antibodies was evaluated in serum samples obtained from 127 household ferrets (Mustela putorius furo) in the Province of Valencia (Spain). Two ferrets tested positive to SARS-CoV-2 (1.57%) by in-house enzyme-linked immunosorbent assay based on receptor binding domain (RBD) of Spike antigen. Furthermore, anti-RBD SARS-CoV-2 antibodies persisted at detectable levels in a seropositive SARS-CoV-2 domestic ferret beyond 129 days since the first time antibodies were detected. This study reports for the first time the evidence of household pet ferrets exposure to SARS-CoV-2 in Spain to date.

scholarly journals Seroprevalence of feline foamy virus in domestic cats in Poland

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Magdalena Materniak-Kornas ◽  
Tadeusz Frymus ◽  
Martin Löchelt ◽  
Jacek Kuźmak
Keyword(s):  
Clinical Symptoms ◽  
Foamy Virus ◽  
Elisa Test ◽  
Domestic Cats ◽  
High Seroprevalence ◽  
Glutathione S Transferase ◽  
Serum Samples ◽  
Immunoblotting Assay ◽  
Adult Cats ◽  
First Time

Abstract Introduction Feline foamy virus (FFVfca) is widespread and its prevalence in naturally infected domestic cats ranges between 30% and 80% worldwide. The infection is persistent, with a sustained antibody response in FFVfca-positive cats; however to date, no defined disease or clinical symptoms have been proved to be associated with it. The goal of the presented study was to determine the prevalence of FFVfca infection in domestic cats in Poland. Material and Methods A total of 223 serum samples collected from domestic cats were tested with a glutathione S-transferase capture ELISA test to detect antibodies specific to capsid (Gag), accessory (Bet) and envelope (Env) FFVfca antigens. A Western blot test was used to confirm the ELISA results. Results The cut-off value for the Gag antigen was established by calculation and evaluation with the immunoblotting assay. The cut-off values for Bet and Env were calculated from the reactivity of Gag-negative samples. The sera of 99 cats (44%) showed reactivity to Gag, those of 80 did so (35.9 %) to Bet, while only 56 samples (25%) were reactive to Env. Only 51 (22.9%) sera were positive for all antigens. The main diagnostic antigen was selected to be Gag. A statistically significant association was found between FFVfca status and the age of the cat. Conclusions This study proved the high seroprevalence of FFVfca in domestic cats in Poland for the first time and confirmed that adult cats are at higher FFVfca infection risk than preadult cats. Its results correspond to those reported from other countries.

scholarly journals Schistosoma haematobium Extracellular Vesicle Proteins Confer Protection in a Heterologous Model of Schistosomiasis

Vaccines ◽  
2020 ◽  
Vol 8 (3) ◽  
pp. 416 ◽  
Author(s):  
Gebeyaw G. Mekonnen ◽  
Bemnet A. Tedla ◽  
Darren Pickering ◽  
Luke Becker ◽  
Lei Wang ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Schistosoma Haematobium ◽  
Extracellular Vesicle ◽  
Surface Proteins ◽  
Helminth Parasites ◽  
Glutathione S Transferase ◽  
Heterologous Challenge ◽  
Host Parasite ◽  
Host Tissues ◽  
Vesicle Proteins

Helminth parasites release extracellular vesicles which interact with the surrounding host tissues, mediating host–parasite communication and other fundamental processes of parasitism. As such, vesicle proteins present attractive targets for the development of novel intervention strategies to control these parasites and the diseases they cause. Herein, we describe the first proteomic analysis by LC-MS/MS of two types of extracellular vesicles (exosome-like, 120 k pellet vesicles and microvesicle-like, 15 k pellet vesicles) from adult Schistosoma haematobium worms. A total of 57 and 330 proteins were identified in the 120 k pellet vesicles and larger 15 k pellet vesicles, respectively, and some of the most abundant molecules included homologues of known helminth vaccine and diagnostic candidates such as Sm-TSP2, Sm23, glutathione S-transferase, saponins and aminopeptidases. Tetraspanins were highly represented in the analysis and found in both vesicle types. Vaccination of mice with recombinant versions of three of these tetraspanins induced protection in a heterologous challenge (S. mansoni) model of infection, resulting in significant reductions (averaged across two independent trials) in liver (47%, 38% and 41%) and intestinal (47%, 45% and 41%) egg burdens. These findings offer insight into the mechanisms by which anti-tetraspanin antibodies confer protection and highlight the potential that extracellular vesicle surface proteins offer as anti-helminth vaccines.

scholarly journals Host-Parasite Relationships in Porcine Ascariosis: Anticoagulant Potential of the Third Larval Stage of Ascaris suum as a Possible Survival Mechanism

Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 804
Author(s):  
Alicia Diosdado ◽  
Fernando Simón ◽  
Rodrigo Morchón ◽  
Javier González-Miguel
Keyword(s):  
Ascaris Suum ◽  
Coagulation Factor ◽  
Coagulation Cascade ◽  
Helminth Parasites ◽  
Factor X ◽  
Larval Stages ◽  
Third Stage ◽  
Migration Strategies ◽  
Host Parasite ◽  
First Time

In order to evade the response of their hosts, helminth parasites have evolved precise and highly regulated mechanisms, including migration strategies of the larval stages. In regard to porcine ascariosis caused by Ascaris suum, its infective third-stage larvae (AsL3) undergo a complex migratory route through the bloodstream of their host before establishing in the small intestine to reach maturation. Despite the benefits attributed to this migration, blood clots formation could compromise larvae survival. The aim of this work was to study the interaction between the cuticle and excretory/secretory antigens of AsL3 and the host coagulation cascade. Larvae were obtained after incubating and hatching A. suum eggs, after which the antigenic extracts were produced. Their ability to disrupt the coagulation cascade was studied using anticoagulation and chromogenic assays, and techniques based on electrophoresis. The obtained results showed that both antigenic extracts possessed anticoagulant potential, being able to inhibit the intrinsic, extrinsic and/or common pathways of the blood coagulation cascade as well as the activated factor X. Moreover, three A. suum serpin proteins were identified as candidates to inhibit this host coagulation factor. To the best of our knowledge, this study shows, for the first time, the anticoagulant potential of the infective larvae of A. suum, which could be used by the parasite as a mechanism to facilitate its invasion and survival in the host.

scholarly journals A Deep Exploration of the Transcriptome and “Excretory/Secretory” Proteome of Adult Fascioloides magna

2012 ◽  
Vol 11 (11) ◽  
pp. 1340-1353 ◽  
Author(s):  
Cinzia Cantacessi ◽  
Jason Mulvenna ◽  
Neil D. Young ◽  
Martin Kasny ◽  
Petr Horak ◽  
...  
Keyword(s):  
Biological Pathway ◽  
Fascioloides Magna ◽  
Protein Motifs ◽  
Sds Page ◽  
Antioxidant Proteins ◽  
Secretory Products ◽  
Liver Flukes ◽  
Host Parasite ◽  
Deep Exploration ◽  
First Time

Parasitic liver flukes of the family Fasciolidae are responsible for major socioeconomic losses worldwide. However, at present, knowledge of the fundamental molecular biology of these organisms is scant. Here, we characterize, for the first time, the transcriptome and secreted proteome of the adult stage of the “giant liver fluke,” Fascioloides magna, using Illumina sequencing technology and one-dimensional SDS-PAGE and OFFGEL protein electrophoresis, respectively. A total of ∼54,000,000 reads were generated and assembled into ∼39,000 contiguous sequences (contigs); ∼20,000 peptides were predicted and classified based on homology searches, protein motifs, gene ontology, and biological pathway mapping. From the predicted proteome, 48.1% of proteins could be assigned to 384 biological pathway terms, including “spliceosome,” “RNA transport,” and “endocytosis.” Putative proteins involved in amino acid degradation were most abundant. Of the 835 secreted proteins predicted from the transcriptome of F. magna, 80 were identified in the excretory/secretory products from this parasite. Highly represented were antioxidant proteins, followed by peptidases (particularly cathepsins) and proteins involved in carbohydrate metabolism. The integration of transcriptomic and proteomic datasets generated herein sets the scene for future studies aimed at exploring the potential role(s) that molecules might play at the host–parasite interface and for establishing novel strategies for the treatment or control of parasitic fluke infections.

Excretory–secretory products of helminth parasites: effects on host immune responses

Parasitology ◽  
1988 ◽  
Vol 96 (S1) ◽  
pp. S123-S166 ◽  
Author(s):  
M. W. Lightowlers ◽  
M. D. Rickard
Keyword(s):  
Immune Responses ◽  
Inflammatory Responses ◽  
Helminth Parasites ◽  
Host Immune Responses ◽  
Parasitic Helminths ◽  
Protective Antigens ◽  
Helminth Infections ◽  
Secretory Products ◽  
Antigen Presenting ◽  
Host Parasite

SUMMARYParasitic helminths excrete or secrete (ES) a variety of molecules into their mammalian hosts. The effects of these ES products on the host's immune responses are reviewed. Investigations into the source of antigenic or immunoregulatory ES products have identified the cuticular and tegumental surfaces of some nematodes and trematodes respectively as being important sources of ES products; other ES molecules are released through specialized excretory or secretory organs. It is proposed that the active shedding of surface antigens may serve as an important source of parasite antigens available to the immune system in a form in which they can be taken up and processed by antigen-presenting dendritic cells, macro-phages and certain B cells for presentation to T helper cells. The ES products of nematodes, trematodes and cestodes contribute to immune evasion strategies of the parasites through mechanisms including shedding of surface-bound ligands and cells, alteration of lymphocyte, macrophage and granulocyte functions and modulation of complement and other host inflammatory responses. Immunopathology may be induced by ES products as in the development of granulomas around entrapped schistosome eggs. In some host – parasite systems ES antigens may induce host-protective immune responses and this source of protective antigens has been utilized in the successful vaccination against helminth infections, particularly against infection with trichurid nematodes and the metacestode stage of cestode parasites. The use of ES antigens in immunodiagnosis of helminth infection is also briefly discussed.

scholarly journals Parasites of South African wildlife. XIX. The prevalence of helminths in some common antelopes, warthogs and a bushpig in the Limpopo province, South Africa

2011 ◽  
Vol 78 (1) ◽  
Author(s):  
Ilana C. Van Wyk ◽  
Joop Boomker
Keyword(s):  
South African ◽  
Fasciola Hepatica ◽  
Limpopo Province ◽  
Helminth Parasites ◽  
Gastrointestinal Helminths ◽  
New Host ◽  
Black Wildebeest ◽  
The Mean ◽  
Connochaetes Taurinus ◽  
Host Parasite

Little work has been conducted on the helminth parasites of artiodactylids in the northern and western parts of the Limpopo province, which is considerably drier than the rest of the province. The aim of this study was to determine the kinds and numbers of helminth that occur in different wildlife hosts in the area as well as whether any zoonotic helminths were present. Ten impalas (Aepyceros melampus), eight kudus (Tragelaphus strepsiceros), four blue wildebeest (Connochaetes taurinus), two black wildebeest (Connochaetes gnou), three gemsbok (Oryx gazella), one nyala (Tragelaphus angasii), one bushbuck (Tragelaphus scriptus), one waterbuck (Kobus ellipsiprymnus), six warthogs (Phacochoerus aethiopicus) and a single bushpig (Potamochoerus porcus) were sampled from various localities in the semi-arid northern and western areas of the Limpopo province. New host–parasite associations included Trichostrongylus deflexus from blue wildebeest, Agriostomum gorgonis from black wildebeest, Stilesia globipunctata from the waterbuck and Fasciola hepatica in a kudu. The mean helminth burden, including extra-gastrointestinal helminths, was 592 in impalas, 407 in kudus and blue wildebeest, 588 in black wildebeest, 184 in gemsbok, and 2150 in the waterbuck. Excluding Probstmayria vivipara, the mean helminth burden in warthogs was 2228 and the total nematode burden in the bushpig was 80. The total burdens and species richness of the helminths in this study were consistently low when compared with similar studies on the same species in areas with higher rainfall. This has practical implications when animals are translocated to areas with higher rainfall and higher prevalence of helminths.

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