A proteomic approach to investigate the differential antigenic profile of two Coxiella burnetii strains

2011 ◽  
Vol 74 (7) ◽  
pp. 1150-1159 ◽  
Author(s):  
Anastasia Papadioti ◽  
Stavroula Markoutsa ◽  
Iosif Vranakis ◽  
Yiannis Tselentis ◽  
Michael Karas ◽  
...  
Author(s):  
K. Cruz ◽  
A. Marcilla ◽  
P. Kelly ◽  
M. Vandenplas ◽  
A. Osuna ◽  
...  

Abstract Background: Trichuris trichiura embryonated eggs are the infectious developmental stage and the first signal to the immune system of the definitive host. Each infective T. trichiura egg carries the antigens needed to challenge the immune system with a wide variety of proteins present in the shell, larvae’s surface, and the accompanying fluid that contains their excretions/secretions. The parasite eggs constitute the first antigenic stimuli to evoke the host response to this intestinal parasite with direct life cycle and enteric development.Methods: The soluble egg extract of T. trichiura obtained from naturally infected African green monkeys (Chlorocebus sabaeus) was investigated using a proteomic approach by mass spectrometry. The antigenic profile of the egg soluble proteins against sera IgG from C. sabaeus with trichuriasis was also investigated by Western blot and LC-MS/MS from the corresponding SDS-PAGE gel.Results: A total of 231 proteins were accurately identified, 168 with known molecular functions. The proteome of the egg lysate revealed common protein families including energy and metabolism; cytoskeleton, motility and muscle; proteolysis; signaling; stress and detoxification; transcription and translation and; lipid binding and transport. Vitellogenin N and VWD and DUF1943 domain containing protein, Poly-cysteine and histidine tailed protein isoform 2, Heat shock protein 70, Glyceraldehyde-3-phosphate dehydrogenase, Actin and Enolase, were among the potential immunoactive proteins. Conclusions: To our knowledge, this study represents the first attempt to identify the proteome of the T. trichiura egg extract as a novel source of immunomodulators and targets for immunodiagnosis able to contribute to the treatment of human autoimmune diseases and to the control of this neglected disease.


2021 ◽  
Vol 15 (3) ◽  
pp. e0009221
Author(s):  
Katalina Cruz ◽  
Antonio Marcilla ◽  
Patrick Kelly ◽  
Michel Vandenplas ◽  
Antonio Osuna ◽  
...  

Embryonated eggs are the infectious developmental stage of Trichuris trichiura and are the primary stimulus for the immune system of the definitive host. The intestinal-dwelling T. trichiura affects an estimated 465 million people worldwide with an estimated global burden of disease of 640 000 DALYs (Disability Adjusted Life Years). In Latin America and the Caribbean, trichuriasis is the most prevalent soil transmitted helminthiasis in the region (12.3%; 95% CI). The adverse health consequences impair childhood school performance and reduce school attendance resulting in lower future wage-earning capacity. The accumulation of the long-term effects translates into poverty promoting sequelae and a cycle of impoverishment. Each infective T. trichiura egg carries the antigens needed to face the immune system with a wide variety of proteins present in the shell, larvae’s surface, and the accompanying fluid that contains their excretions/secretions. We used a proteomic approach with tandem mass spectrometry to investigate the proteome of soluble non-embryonated egg extracts of T. trichiura obtained from naturally infected African green monkeys (Chlorocebus sabaeus). A total of 231 proteins were identified, 168 of them with known molecular functions. The proteome revealed common proteins families which are known to play roles in energy and metabolism; the cytoskeleton, muscle and motility; proteolysis; signaling; the stress response and detoxification; transcription and translation; and lipid binding and transport. In addition to the study of the T. trichiura non-embryonated egg proteome, the antigenic profile of the T. trichiura non-embryonated egg and female soluble proteins against serum antibodies from C. sabaeus naturally infected with trichuriasis was investigated. We used an immunoproteomic approach by Western blot and tandem mass spectrometry from the corresponding SDS-PAGE gels. Vitellogenin N and VWD and DUF1943 domain containing protein, poly-cysteine and histidine tailed protein isoform 2, heat shock protein 70, glyceraldehyde-3-phosphate dehydrogenase, actin, and enolase, were among the potential immunoactive proteins. To our knowledge, this is the first study on the T. trichiura non-embryonated egg proteome as a novel source of information on potential targets for immunodiagnostics and immunomodulators from a neglected tropical disease. This initial list of T. trichiura non-embryonated egg proteins (proteome and antigenic profile) can be used in future research on the immunobiology and pathogenesis of human trichuriasis and the treatment of human intestinal immune-related diseases.


Author(s):  
T. F. McCaul ◽  
R. J. Gould

Immuno-electron microscopy has allowed the selective localisation of molecules with high resolution and high specificity. Cryopreparatory methods have provided better retention of antigenicity suitable for precise immunolabelling together with optimal structural preservation of cellular components. Cryosubstitution and cryoultramicrotomy have widely been exploited for immunolabelling. Molecular Distillation Dryer (MDD), a form of freeze-drying technique, has recently been used for immunolabelling of Plasmodium falciparum stress proteins and nuclear ribonucleoprotein particles in cultured cells. In the present study, we report the comparison of all three cryotechniques in the immunolabelling of bacterial antigens of Coxiella burnetii.The highly infectious C. burnetii was prefixed in 3% glutaraldehyde (66 mM cacodylate buffer, pH 6.8 ). The cells were then pre-embedded in 2% low-temperature agarose on Durapore hydrophilic membrane prior to cryofixation using a LifeCell CF100 metal-mirror system. A 1% glutaraldehyde in 100% methanol was used as a medium for cryosubstitution in a Reichert CS Auto Cryosubstitution apparatus.


2010 ◽  
Vol 72 (08/09) ◽  
Author(s):  
H Bernard ◽  
S Brockmann ◽  
N Kleinkauf ◽  
C Klinc ◽  
C Wagner-Wiening ◽  
...  

2014 ◽  
Author(s):  
Sung-Jae Yoon ◽  
Woo-Sung Kwon ◽  
Saidur Rahman ◽  
June-Sub Lee ◽  
Yoo-Jin Park ◽  
...  

2003 ◽  
Author(s):  
Charles Thomas Parker ◽  
Dorothea Taylor ◽  
George M Garrity
Keyword(s):  

2015 ◽  
Vol 12 (1) ◽  
pp. 1 ◽  
Author(s):  
Rosmilah Misnan ◽  
Nurul Izzah Abdul Rahman ◽  
Zailatul Hani Mohd Yadzir ◽  
Noormalin Abdullah ◽  
Mohd Faizal Bakhtiar ◽  
...  

Crab meat is widely consumed in several countries around the world. However, when consumed, crab meats are frequent cause of allergic reactions throughout the world. Scylla serrata is among the most common mud crab in Malaysia. In a previous study two major allergens of mud crab at 36 and 41 kDa was identified. Thus, the aim of this study is to further identify these major allergens by a proteomic approach. Protein extract was prepared and resolved by 2-dimensional electrophoresis (2-DE). Immunoblotting was then performed using reactive sera from patients with crab allergy. Major allergenic spots were then excised from the 2-DE gel and analysed by mass spectrometry. The 2-DE profile of the extract revealed approximately >100 protein spots between pH of 4.00 to 8.00. Mass spectrometry analysis has identified the 36 and 41 kDa proteins as tropomyosin and arginine kinase, respectively. Our findings indicated that tropomyosin and arginine kinase play a major role in allergic reaction to mud crab meat among local patients with crab meat allergy, and should be included in diagnostics and therapeutic strategies of this allergy.


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