scholarly journals Leucine Aminopeptidase Is an Immunodominant Antigen of Fasciola hepatica Excretory and Secretory Products in Human Infections

2007 ◽  
Vol 15 (1) ◽  
pp. 95-100 ◽  
Author(s):  
A. Marcilla ◽  
J. E. De la Rubia ◽  
J. Sotillo ◽  
D. Bernal ◽  
C. Carmona ◽  
...  
Keyword(s):  
Phosphoenolpyruvate Carboxykinase ◽  
Leucine Aminopeptidase ◽  
Fasciola Hepatica ◽  
Enzyme Linked Immunosorbent Assay ◽  
Strong Reaction ◽  
Immunodominant Antigen ◽  
Proteomic Approach ◽  
Immunodominant Antigens ◽  
Secretory Products ◽  
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 recombinant LAP as the antigen was developed to check sera from individuals of this region. Our results demonstrate that LAP produces a specific and strong reaction, suggesting its potential use in the serologic diagnosis of F. hepatica infections in humans.

Excretory–secretory products of larval Fasciola hepatica investigated using a two-dimensional proteomic approach

2008 ◽  
Vol 161 (1) ◽  
pp. 63-66 ◽  
Author(s):  
Benjamin E.F. Gourbal ◽  
François Guillou ◽  
Guillaume Mitta ◽  
Pierre Sibille ◽  
André Thèron ◽  
...  
Keyword(s):  
Fasciola Hepatica ◽  
Two Dimensional ◽  
Proteomic Approach ◽  
Secretory Products

scholarly journals Performance of a 27 kDa Fasciola hepatica Antigen in the Diagnosis of Human Fascioliasis

2015 ◽  
Vol 7 (01) ◽  
pp. 017-020 ◽  
Author(s):  
Reza Shafiei ◽  
Bahador Sarkari ◽  
Seyed Mahmoud Sadjjadi
Keyword(s):  
Sensitivity And Specificity ◽  
Western Blotting ◽  
Indirect Elisa ◽  
Fasciola Hepatica ◽  
Serological Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Controls ◽  
Elisa System ◽  
Immunodominant Antigen ◽  
Human Fascioliasis

ABSTRACT Background: Serological diagnosis, based on antigenic fractions of the parasite can be used for the early diagnosis of human fascioliasis. The current study aimed to evaluate the efficacy of a 27 kDa immunodominant antigen of Fasciola hepatica adult worms, in an indirect enzyme-linked immunosorbent assay (ELISA) system for serological diagnosis of human fascioliasis. Materials and Methods: The immunodiagnosis of human fascioliasis, using a 27 kDa immunodominant antigen, purified from F. hepatica somatic antigens (SAs), was evaluated by Western blotting and ELISA with sera samples of human fascioliasis patients, healthy controls and patients with other parasitic infections. Results: Using western blotting, from 12 sera of fascioliasis patients, 11 sera (91.6%) detected the 27 kDa subunit. None of 30 samples from healthy controls or 32 sera from nonfascioliasis patients reacted with the 27 kDa antigen. Accordingly, sensitivity and specificity of the system was found to be 91.6% and 100%, respectively. The 27 kDa antigen was purified from the SAs and was used in an indirect ELISA system. Of 15 sera of fascioliasis patients, all (100%) were found to be positive by ELISA whereas only 4 cases (6.25%) of nonfascioliasis patients or healthy controls were false-positive by this system. Accordingly, the sensitivity and specificity of the test were 100% and 93.6%, respectively. Conclusion: Findings of this study demonstrated that both Western blotting and the indirect ELISA, based on the 27 kDa subunit of F. hepatica SA, are reliable methods for serodiagnosis of human fascioliasis.

scholarly journals Evaluation and Characterization of Fasciola hepatica Tegument Protein Extract for Serodiagnosis of Human Fascioliasis

2012 ◽  
Vol 19 (11) ◽  
pp. 1870-1878 ◽  
Author(s):  
Adelaida Morales ◽  
Ana M. Espino
Keyword(s):  
Fasciola Hepatica ◽  
Tegument Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Parasitic Infections ◽  
Protein Extract ◽  
Fatty Acid Binding ◽  
Content Type ◽  
Proteomic Approach ◽  
Human Fascioliasis ◽  
Diagnostic Agent

ABSTRACTTegument protein extract fromFasciola hepaticaadult flukes (FhTA) was obtained and assessed for its potential as a diagnostic agent for the serological detection of human fascioliasis using an indirect enzyme-linked immunosorbent assay (ELISA). In an analysis of sera from 45 patients infected withF. hepatica, sera from 41 patients with other parasitic infections, and sera from 33 healthy controls, the FhTA-ELISA showed sensitivity, specificity, and accuracy of 91.1%, 97.3%, and 95%, respectively. Specific IgG1 and IgG4 were the antibody isotypes mainly detected in sera from patients with fascioliasis. Polypeptides of 52, 38, 24 to 26, and 12 to 14 kDa were identified by Western blotting as the most immunoreactive components of the FhTA. A proteomic approach led us to identify enolase, aldolase, glutathioneS-transferase, and fatty acid binding protein as the major immunoreactive components of the FhTA.

Antigenic Components of Excretory-Secretory Products of Adult Fasciola hepatica Recognized in Human Infections

1996 ◽  
Vol 54 (2) ◽  
pp. 146-148 ◽  
Author(s):  
M. L. Sampaio Silva ◽  
L. Monjour ◽  
J. M. Correia da Costa ◽  
M. A. Pires ◽  
A. M. Viana da Costa ◽  
...  
Keyword(s):  
Fasciola Hepatica ◽  
Secretory Products ◽  
Human Infections

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 Characterization of the Humoral Immune Response to Chlamydia Outer Membrane Protein 2 in Chlamydial Infection

2003 ◽  
Vol 10 (1) ◽  
pp. 103-107 ◽  
Author(s):  
I. Portig ◽  
J. C. Goodall ◽  
R. L. Bailey ◽  
J. S. H. Gaston
Keyword(s):  
Membrane Protein ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
Chlamydial Infection ◽  
Chlamydia Pneumoniae ◽  
Specific Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Recent Infection ◽  
Humoral Immune ◽  
Immunodominant Antigen

ABSTRACT Detection of antibodies to an outer membrane protein 2 (OMP2) by enzyme-linked immunosorbent assay (ELISA) by using either the Chlamydia trachomatis- or the Chlamydia pneumoniae-specific protein was investigated. OMP2 is an immunodominant antigen giving rise to antibody responses in humans infected with different C. trachomatis serovars (A to C and D to K) or with C. pneumoniae, which could be detected by OMP2 ELISA. OMP2 ELISA is not species specific, but antibody titers were usually higher on the homologous protein. The sensitivity of this assay was high but varied according to the “gold standard” applied. Levels of antibody to C. pneumoniae OMP2 as detected by ELISA seem to return to background or near-background values within a shorter period of time compared to antibodies to C. pneumoniae detected by microimmunofluorescence (MIF), making it more likely that positive results in ELISA reflect recent infection. Thus, OMP2 ELISA has distinct advantages over MIF and commercially available ELISAs and might be a useful tool for the serodiagnosis of chlamydial infection.

Partial Characterization of the Epitope on Excretory-Secretory Products of Fasciola hepatica Recognized by Monoclonal Antibody ES78

1998 ◽  
Vol 84 (1) ◽  
pp. 55 ◽  
Author(s):  
Ailen Diaz ◽  
Ana M. Espino ◽  
Ricardo Marcet ◽  
Oscar Otero ◽  
Diamela Torres ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Fasciola Hepatica ◽  
Partial Characterization ◽  
Secretory Products

Monoclonal antibodies multireactive with parasite antigens produced by hybridomas generated from naive mice

Parasitology ◽  
1997 ◽  
Vol 115 (4) ◽  
pp. 387-393 ◽  
Author(s):  
E. KONISHI
Keyword(s):  
Monoclonal Antibodies ◽  
Fasciola Hepatica ◽  
Toxocara Canis ◽  
Plasmodium Yoelii ◽  
Natural Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Soluble Antigen ◽  
Lethal Challenge ◽  
Multiple Antigen

Hybridoma clones producing IgM naturally occurring (natural) antibodies were generated from naive Balb/c mice and characterized for reactivity against parasite antigens. Ascites of mice injected with these hybridomas reacted with Toxoplasma gondii soluble antigen at levels approximately 1000–10000 times higher than serum pooled from mice used for generating hybridomas as determined by a conventional enzyme-linked immunosorbent assay. Western blot analysis indicated that these monoclonal antibodies reacted with multiple antigen molecules of T. gondii with patterns similar to that of pooled mouse sera. These antibodies also reacted with multiple antigens of Plasmodium yoelii, Entamoeba histolytica, Ascaris lumbricoides, Toxocara canis, Trichuris vulpis, Fasciola hepatica, Schistosoma mansoni, and Dipylidium caninum. When one monoclonal antibody was absorbed with these parasite antigens, its reactivity with T. gondii antigen molecules was consistently reduced, independent of parasite species. These natural antibodies failed to kill T. gondii tachyzoites in vitro or to protect mice from lethal challenge with T. gondii. These results indicate that natural antibodies detected in sera of naive mice are secreted from certain B cell populations and that these antibodies are multireactive with parasite antigens and have low affinity.

scholarly journals A proteomic approach to understand the clinical significance of acute myeloid leukemia-derived extracellular vesicles reflecting essential characteristics of leukemia

2020 ◽  
pp. mcp.RA120.002169
Author(s):  
Ka-Won Kang ◽  
Hyoseon Kim ◽  
Woojune Hur ◽  
Jik-han Jung ◽  
Su Jin Jeong ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Cell Lines ◽  
Myeloid Leukemia ◽  
Extracellular Vesicle ◽  
The Cancer Genome Atlas ◽  
Enzyme Linked Immunosorbent Assay ◽  
Expression Levels ◽  
Proteomic Approach ◽  
Cancer Genome Atlas ◽  
Acute Myeloid

Extracellular vesicle (EV) proteins from acute myeloid leukemia (AML) cell lines were analyzed using mass spectrometry. The analyses identified 2450 proteins, including 461 differentially expressed proteins (290 upregulated and 171 downregulated). CD53 and CD47 were upregulated and were selected as candidate biomarkers. The association between survival of patients with AML and the expression levels of CD53 and CD47 at diagnosis was analyzed using mRNA expression data from The Cancer Genome Atlas database. Patients with higher expression levels showed significantly inferior survival than those with lower expression levels. Enzyme-linked immunosorbent assay results of the expression levels of CD53 and CD47 from EVs in the bone marrow of patients with AML at diagnosis and at the time of complete remission with induction chemotherapy revealed that patients with downregulated CD53 and CD47 expression appeared to relapse less frequently. Network model analysis of EV proteins revealed several upregulated kinases, including LYN, CSNK2A1, SYK, CSK, and PTK2B. The potential cytotoxicity of several clinically applicable drugs that inhibit these kinases was tested in AML cell lines. The drugs lowered the viability of AML cells. The collective data suggest that AML-derived EVs could reflect essential leukemia biology.

scholarly journals Synovial Fluid Cell Proteomic Analysis Identifies Upregulation of Alpha-Taxilin Proteins in Rheumatoid Arthritis: A Potential Prognostic Marker

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Ashish Sarkar ◽  
Shivani Sharma ◽  
Prachi Agnihotri ◽  
Tanmoy Sarkar ◽  
Pooja Kumari ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Synovial Fluid ◽  
Prognostic Marker ◽  
Surrounding Tissue ◽  
Enzyme Linked Immunosorbent Assay ◽  
Two Dimensional Gel Electrophoresis ◽  
Flight Mass Spectrometry ◽  
Tissue Identification ◽  
Proteomic Approach ◽  
In Silico Studies

Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory disease affecting the joints and surrounding tissue. Identification of novel proteins associated with the progression of a disease is a prerequisite for understanding the pathogenesis of RA. The present study was undertaken to identify the potential biomarkers from a less explored biological sample such as synovial fluid (SF) cells which is specific for RA and to analyze their functional aspects using proteomic approach. Two-dimensional gel electrophoresis (2-DE) was performed using synovial fluid cells of RA and osteoarthritis (OA) patients, and 7 differentially expressed proteins were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS/MS). Αlpha-Taxilin (α-Taxilin) has been found as one of the novel, significantly up regulated protein in RA. It has been validated in the synovium, synovial fluid (SF), SF cells, and plasma samples by Western blot, enzyme-linked immunosorbent assay (ELISA), fluorescence-activated cell sorting (FACS), immunohistochemistry (IHC), and real-time PCR. The identification of autoantibody against α-Taxilin and in silico studies has further helped us to understand its involvement in disease mechanism. The present study will therefore provide knowledge towards the etiology of RA that pave the way for suitable prognostic marker identification along with other clinical parameters.

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