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 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 FhCaBP4: a Fasciola hepatica calcium-binding protein with EF-hand and dynein light chain domains

2012 ◽  
Vol 111 (4) ◽  
pp. 1707-1713 ◽  
Author(s):  
Rebecca Orr ◽  
Ruth Kinkead ◽  
Richard Newman ◽  
Lindsay Anderson ◽  
Elizabeth M. Hoey ◽  
...  
Keyword(s):  
Light Chain ◽  
Calcium Binding ◽  
Fasciola Hepatica ◽  
Binding Protein ◽  
Calcium Binding Protein ◽  
Dynein Light Chain ◽  
Ef Hand

scholarly journals FhCaBP2: a Fasciola hepatica calcium-binding protein with EF-hand and dynein light chain domains

Parasitology ◽  
2015 ◽  
Vol 142 (11) ◽  
pp. 1375-1386 ◽  
Author(s):  
CHARLOTTE M. THOMAS ◽  
DAVID J. TIMSON
Keyword(s):  
Light Chain ◽  
Calcium Binding ◽  
Fasciola Hepatica ◽  
Biochemical Properties ◽  
Binding Activity ◽  
Ion Binding ◽  
Dynein Light Chain ◽  
Manganese Ions ◽  
Ef Hand ◽  
The One

SUMMARYFhCaBP2 is a Fasciola hepatica protein which belongs to a family of helminth calcium-binding proteins which combine an N-terminal domain containing two EF-hand motifs and a C-terminal dynein light chain-like (DLC-like) domain. Its predicted structure showed two globular domains joined by a flexible linker. Recombinant FhCaBP2 interacted reversibly with calcium and manganese ions, but not with magnesium, barium, strontium, copper (II), colbalt (II), iron (II), nickel, lead or potassium ions. Cadmium (II) ions appeared to bind non-site-specifically and destabilize the protein. Interaction with either calcium or magnesium ions results in a conformational change in which the protein's surface becomes more hydrophobic. The EF-hand domain alone was able to interact with calcium and manganese ions; the DLC-like domain was not. Alteration of a residue (Asp-58 to Ala) in the second EF-hand motif in this domain abolished ion-binding activity. This suggests that the second EF-hand is the one responsible for ion-binding. FhCaBP2 homodimerizes and the extent of dimerization was not affected by calcium ions or by the aspartate to alanine substitution in the second EF-hand. The isolated EF-hand and DLC-like domains are both capable of homodimerization. FhCaBP2 interacted with the calmodulin antagonists trifluoperazine, chlorpromazine, thiamylal and W7. Interestingly, while chlorpromazine and thiamylal interacted with the EF-hand domain (as expected), trifluoperazine and W7 bound to the DLC-like domain. Overall, FhCaBP2 has distinct biochemical properties compared with other members of this protein family from Fasciola hepatica, a fact which supports the hypothesis that these proteins have different physiological roles.

scholarly journals Proteomic Analysis of Fasciola hepatica Excretory and Secretory Products Co-Immunoprecipitated Using Time Course Infection Sera

Pathogens ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 749
Author(s):  
Zhuo Lan ◽  
Xiao-Lei Liu ◽  
Qing-Bo Lv ◽  
Min-Hao Zeng ◽  
Jun-Feng Gao ◽  
...  
Keyword(s):  
Time Course ◽  
Fasciola Hepatica ◽  
Interacting Proteins ◽  
Metabolism Regulation ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass ◽  
Ruminant Animals ◽  
Secretory Products ◽  
Harmful Effects ◽  
Meaningful Data

Fasciola hepatica is a widespread pathogen that is known for its harmful effects on the health and productivity of ruminant animals. To identify the proteins present in all periods of infection with F. hepatica but not in those with Fasciola gigantica by shotgun liquid chromatography–tandem mass spectrometry (LC–MS/MS), we collected the ESPs and sera of F. hepatica and F. gigantica. In this study, the sheep were artificially infected with F. hepatica and the sera were collected at five different periods: 3 days post-infection (dpi), 7 dpi, 21 dpi, 63 dpi, and 112 dpi. The interacting proteins were pulled down from the sheep sera of all five periods and the sera with F. gigantica by co-immunoprecipitation (Co-IP) assay, before being identified by LC–MS/MS analysis. Thirty, twenty-two, twenty-three, twenty-seven, and twenty-two proteins were pulled down by the infected sera at 3 dpi, 7 dpi, 21 dpi, 63 dpi, and 112 dpi, respectively. Among them, 12 proteins existed in all periods, while six proteins could be detected in all periods in F. hepatica but not in F. gigantica. Protein relative pathway analysis revealed that these proteins mainly refer to the metabolism, regulation of genetic activity, and signal transduction of F. hepatica. In conclusion, this study provides meaningful data for the diagnosis of fasciolosis and to understand the interactions between F. hepatica and the host.

scholarly journals Fasciola hepatica calcium-binding protein FhCaBP2: structure of the dynein light chain-like domain

2016 ◽  
Vol 115 (7) ◽  
pp. 2879-2886 ◽  
Author(s):  
Thanh H. Nguyen ◽  
Charlotte M. Thomas ◽  
David J. Timson ◽  
Mark J. van Raaij
Keyword(s):  
Light Chain ◽  
Calcium Binding ◽  
Fasciola Hepatica ◽  
Binding Protein ◽  
Calcium Binding Protein ◽  
Dynein Light Chain

scholarly journals FhCaBP3: A Fasciola hepatica calcium binding protein with EF-hand and dynein light chain domains

Biochimie ◽  
2013 ◽  
Vol 95 (4) ◽  
pp. 751-758 ◽  
Author(s):  
Samantha Banford ◽  
Orla Drysdale ◽  
Elizabeth M. Hoey ◽  
Alan Trudgett ◽  
David J. Timson
Keyword(s):  
Light Chain ◽  
Calcium Binding ◽  
Fasciola Hepatica ◽  
Binding Protein ◽  
Calcium Binding Protein ◽  
Dynein Light Chain ◽  
Ef Hand

Methodical guidelines on the control of fasciolosis in cattle in the Kaluga region

2018 ◽  
Vol 12 (2) ◽  
pp. 108-116
Author(s):  
A.M. Ustinov ◽  
R.T. Safiullin ◽  
R.R. Safiullin
Keyword(s):  
Infected Animal ◽  
Fasciola Hepatica ◽  
Adult Stage ◽  
Federal District ◽  
Dosage Forms ◽  
Pond Snail ◽  
Meat Processing ◽  
Processing Plants ◽  
Causative Agents ◽  
Kaluga Region

Information is provided on the causative agents of fasciolosis and the developmental biology of Fasciola hepatica. Fasciolosis of cattle is widespread in the conditions of the Kaluga Region and the Central Federal District on farms of different forms of ownership. The greatest number of F. hepatica eggs in 1 g of feces in cattle is established in JSC "Iskra" - 175,4 specimens, the lowest - in the samples from cows from private farms - 87,6 sp. At the opening slaughter of cattle at meat processing plants region and slaughterhouses invasion of their fasciolae was 34.8%. The average number fascial one infected animal was 32.1 specimens, including adult - and larval specimens of 25.2 and 6.9 copies At veterinary-sanitary expertise and helminthological autopsy of the liver, gallbladder and bile ducts installed year-round invasion fasciolae mainly in the adult stage. Young animals of the current year are infected with fasciola in the pasture massly in August-September, and the extent of invasion in November was 4.3%, for winter-stabling months an average of 12.6%, in March - 17.9% and in April after de-worming - 5.9%.The data obtained in the study of the distribution of the small pond has shown that in the Kaluga region these mollusks are widespread. In this case, the invasion of the small pond snail by larval forms of fasciolae in different years has its own peculiarities, which are influenced by climatic factors.The different dosage forms of clozantel - Rolenol and Santel and Genamectin with mixed fascioliasis and hypodermatosis of bovine in the recommended dose against subcutaneous gadfly larvae - provided 100% efficacy. The tested dosage forms of clozantel are similar in effectiveness against subcutaneous gadfly larvae (EE-100%) and fasciol (EE-95 and 90%, IE-97.4 and 96.3%).The therapeutic and economic effectiveness of complex domestic preparations of Helmicide and Fascocida in spontaneous fascioliasis of cattle is studied.

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