Cloning, expression, characterization, and immunological properties of citrate synthase from Echinococcus granulosus

Wang, Ning; Zhu, Hui; Zhan, Jiafei; Guo, Cheng; Shen, Nengxing; Gu, Xiaobin; Lai, Weimin; Xie, Yue; Peng, Xuerong; Yang, Guangyou

doi:10.1007/s00436-019-06334-6

Cited by 12 publications

(4 citation statements)

References 47 publications

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“…Whether the high expression of fructosebisphosphate aldolase and enolase is beneficial for parasite's growth and development needs to be investigated in future. Previous studies showed enolase has primarily functioned as glycolytic enzymes in the cytoplasm (13)(14)(15). Besides the cytoplasmic localization, enolase was detected in the tegument of protoscolexes and adult worms, suggesting potentially participated in host-parasite molecular crosstalk (13).…”

Section: Resultsmentioning

confidence: 95%

Proteomics Analysis of Hydatigera taeniaeformis Metacestode Stage

Guo

2020

Front. Vet. Sci.

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Hydatigera taeniaeformis (H. taeniaeformis) is one of the most robust of tapeworm parasites that is widely distributed around the world. Information of proteins of H. taeniaeformis has not previously been reported. Using liquid chromatography tandem-mass spectrometry (LC-MS/MS) analysis, the proteome of H. taeniaeformis metacestode was profiled and a total of 408 proteins were identified. Of these, 26.5% (108/408) were annotated to be associated with metabolic pathways. Consistently, Gene Ontology analysis showed that those identified proteins were mainly classified into metabolic process of the biological processes. A set of metabolic enzymes, including Fructose-1,6-bisphosphate aldolase, enolase, Glucan phosphorylase, and phosphoenolpyruvate carboxykinase, were abundant in H. taeniaeformis metacestodes. In addition, some rare but interesting proteins like antigens (such as tegument protein and Antigen B) were identified. The two recombinant proteins of tegument protein and Antigen B were well-recognized by the sera from the H. taeniaeformis-infected mice. The H. taeniaeformis metacestode proteome might help to find new candidates for the immunodiagnosis and vaccine development and provide valuable information on H. taeniaeformis biology.

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Section: Resultsmentioning

confidence: 95%

Proteomics Analysis of Hydatigera taeniaeformis Metacestode Stage

Guo

2020

Front. Vet. Sci.

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“…ELISA has also been shown to be very efficient because of its high sensitivity as well as the need for small amounts of antigen. Several authors have reported success using this technique with crude antigen preparations (Lightowlers et al, 1989;Wen et al, 2019), purified antigens (De la Rue et al, 2010;Manzano-Roman et al, 2015;Iraqi, 2016) and recombinant antigens (Faramarzi et al, 2019;Wang et al, 2019). Petrone et al (2021) evaluated the accuracy of a wholeblood stimulation test based on interleukin-4 and other immune/growth factors detection in response to AgB of E. granulosus sensu lato, to discriminate cyst viability and detect cyst reactivation in patients with hepatic CE.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of two heterologous recombinant antigens for the serological diagnosis of human polycystic echinococcosis

et al. 2022

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Polycystic echinococcosis (PE) is a zoonosis endemic in the Neotropical region of the Americas. It is caused by the larval stage of the cestode Echinococcus vogeli, which develops as harmful cysts that slowly grow in the liver, lungs and other organs of humans and other host species. Human PE diagnosis is usually based on clinical and epidemiological aspects and imaging techniques, often requiring confirmation by immunological assays. The currently available serological tests for detecting antibodies against Echinococcus spp. are mostly based on complex, variable and poorly characterized mixtures of native parasite antigens, which impairs specificity and/or sensitivity. In this scenario, the evaluation of well-characterized alternative antigens is urgently needed for the improvement of PE diagnosis. Here, two subunits (AgB8/1 and AgB8/2) of the major secretory antigen from Echinococcus granulosus (antigen B (AgB)), of diagnostic value for cystic echinococcosis, were validated for PE diagnosis. These antigens, produced as pure recombinant proteins (rAgB8/1 and rAgB8/2) in Escherichia coli, allowed detecting specific immunoglobulin G antibodies in sera from PE patients in an enzyme-linked immunosorbent assay, with sensitivities of 83.72% and 81.40%, respectively, and specificities of 83.12% and 80.09%, respectively. The use of recombinant proteins overcomes difficulties to obtain parasite material and reduced non-specific reactions and costs. Our results demonstrated reproducibility and accuracy high enough to be considered valid according to the acceptance criteria for Food and Drug Administration assay validation. This qualifies rAgB8/1 and rAgB8/2 as potential substitutes for the currently used parasite crude or partially purified antigens.

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“…Furthermore, the location of EGR-03347 in the suction cups and hooks of protocercus might be related to the energy demand attached to the host intestine, thus EGR-03347 might be involved in the interaction between the parasite and its host [20,21]. This suggested that EGR-03347 plays a key role in the growth and physiological activities of E. granulosus [22]. Tapeworms contain the complete metabolic pathways of fat, which could be used for the formation and development of the ascus and PSC [23,24].…”

Section: Discussionmentioning

confidence: 99%

Molecular Characterization and Expression Analysis of the Gene Encoding 3-Hydroxyacyl-CoA Dehydrogenase (EGR-03347) from Echinococcus Granulosus and the Evaluation of the Immune Protection of the Definitive Hosts (dogs)

Xian¹,

Wang²,

Zhao³

et al. 2021

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Background: Cystic echinococcosis, a serious parasitic zoonosis, is caused by tapeworm (Echinococcus granulosus) larvae. The development of an effective vaccine is a promising strategy to control echinococcosis. E. granulosus has a complete tricarboxylic acid cycle pathway, in which 3-hydroxyacyl-CoA dehydrogenase (EGR-03347) is a key enzyme.Methods: In the present study, the cDNA encoding EGR-03347 in Echinococcus granulosus (rEGR-03347) was successfully cloned and the molecular and biochemical characterizations carried out. The immunoreactivity of recombinant EGR-03347 (rEGR-03347) was investigated using western blotting. The immunolocalization of EGR-03347 in different life stages of E. granulosus was determined using specific polyclonal antibody, quantitative real-time reverse transcription polymerase chain reaction was used to analyze their transcript levels in PSCs and 28-day strobilated worms stages. In addition, recombinant protein rEGR-03347 was mixed with the adjuvant Quil A for vaccinating dogs, after three vaccine injections, all the dogs were orally challenge-infected with 100000 protoscoleces of E. granulosus. After 28 days of infection, all the dogs were euthanized and necropsied for collecting and counting E. granulosus worms, post-infection the antibody and cytokine were measured for the immunogenicity analysis of this protein.Results: rEGR-03347 is a highly conserved protein, consisting of 308 amino acids. Recombinant EGR-03347 could be identifed in the sera of patients with CE and in mouse anti-rEGR-03347 sera. Immunofluorescence analysis showed that EGR-03347 mainly localized in the tegument of protoscoleces and adults, and their transcript levels were high in the 28-day strobilated worms. Furthermore, enzyme-linked immunosorbent assays post‑infection showed that IgG gradually increased after the first immunization with rEGR-03347 compared with the control group, rEGR-03347 changed the interferon gamma and interleukin 4 levels. We observed an 87.2 % reduction in E. granulosus numbers and 66.7 % inhibition of the segmental development of E. granulosus in the rEGR‑03347‑vaccinated dogs compared with the nonvaccinated controls.Conclusions: This is the first report characterizing a 3-hydroxyacyl-CoA dehydrogenase from the tapeworm E. granulosus. We have characterized the sequence, structure and location of EGR‑03347 and investigated the immunoreactivity, immunogenicity and serodiagnostic potential of rEGR‑03347 . The results demonstrated rEGR-03347 as a potential vaccine against E. granulosus infection in dogs.

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Cloning, expression, characterization, and immunological properties of citrate synthase from Echinococcus granulosus

Cited by 12 publications

References 47 publications

Proteomics Analysis of Hydatigera taeniaeformis Metacestode Stage

Proteomics Analysis of Hydatigera taeniaeformis Metacestode Stage

Evaluation of two heterologous recombinant antigens for the serological diagnosis of human polycystic echinococcosis

Molecular Characterization and Expression Analysis of the Gene Encoding 3-Hydroxyacyl-CoA Dehydrogenase (EGR-03347) from Echinococcus Granulosus and the Evaluation of the Immune Protection of the Definitive Hosts (dogs)

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