A recombinant thioredoxin-glutathione reductase from Fasciola hepatica induces a protective response in rabbits

Maggioli, Gabriela; Silveira, Fernando; Martı́n-Alonso, José M.; Salinas, Gustavo; Carmona, Carlos; Parra, Francisco

doi:10.1016/j.exppara.2011.09.013

Cited by 29 publications

(9 citation statements)

References 37 publications

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“…The construct for recombinant expression of E. granulosus TGR was previously generated [15].The construct for F. hepatica TGR was generated using the same methodology [18]. Both TGR constructs contained the Sec insertion sequence (SECIS) element of E. coli formate dehydrogenase H at a 10 nucleotide distance from the penultimate UGA Sec codon, to allow stop codon recoding to Sec, as previously described [15], [18].…”

Section: Methodsmentioning

confidence: 99%

“…Both TGR constructs contained the Sec insertion sequence (SECIS) element of E. coli formate dehydrogenase H at a 10 nucleotide distance from the penultimate UGA Sec codon, to allow stop codon recoding to Sec, as previously described [15], [18]. For recombinant protein expression, TGR constructs were used to transform E. coli BL21(DE3) cells previously transformed with pSUABC, a plasmid that encodes selA , selB and selC , supporting high level expression of the genes involved in Sec incorporation [19].…”

Section: Methodsmentioning

confidence: 99%

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Identification of Thioredoxin Glutathione Reductase Inhibitors That Kill Cestode and Trematode Parasites

et al. 2012

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Parasitic flatworms are responsible for serious infectious diseases that affect humans as well as livestock animals in vast regions of the world. Yet, the drug armamentarium available for treatment of these infections is limited: praziquantel is the single drug currently available for 200 million people infected with Schistosoma spp. and there is justified concern about emergence of drug resistance. Thioredoxin glutathione reductase (TGR) is an essential core enzyme for redox homeostasis in flatworm parasites. In this work, we searched for flatworm TGR inhibitors testing compounds belonging to various families known to inhibit thioredoxin reductase or TGR and also additional electrophilic compounds. Several furoxans and one thiadiazole potently inhibited TGRs from both classes of parasitic flatworms: cestoda (tapeworms) and trematoda (flukes), while several benzofuroxans and a quinoxaline moderately inhibited TGRs. Remarkably, five active compounds from diverse families possessed a phenylsulfonyl group, strongly suggesting that this moiety is a new pharmacophore. The most active inhibitors were further characterized and displayed slow and nearly irreversible binding to TGR. These compounds efficiently killed Echinococcus granulosus larval worms and Fasciola hepatica newly excysted juveniles in vitro at a 20 µM concentration. Our results support the concept that the redox metabolism of flatworm parasites is precarious and particularly susceptible to destabilization, show that furoxans can be used to target both flukes and tapeworms, and identified phenylsulfonyl as a new drug-hit moiety for both classes of flatworm parasites.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Identification of Thioredoxin Glutathione Reductase Inhibitors That Kill Cestode and Trematode Parasites

et al. 2012

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“…Over the last decade a large number of native and recombinant parasite proteins have been identified and tested as vaccine in a number of animals models. Some of these molecules includes leucinaminopeptidase (Maggioli et al, 2011a), cathepsin-L (Chantree et al, 2013; Golden et al, 2010; Piacenza et al, 1999), thioredoxin-glutathion reductase (Maggioli et al, 2011b), glutathione S-transferase (Sexton et al, 1990; Sexton et al, 1994), fatty acid binding protein (Casanueva et al, 2001; Martinez-Fernandez et al, 2004) and saposin-like protein-2 (Espino et al, 2010; Kueakhai et al, 2013). All these vaccine candidates have induced partial levels of protection that range between 46.9–83% in different animal models, which indicates that a vaccine against F. hepatica is not a chimera but an attainable goal.…”

Section: Introductionmentioning

confidence: 99%

Adjuvant-enhanced antibody and cellular responses to inclusion bodies expressing FhSAP2 correlates with protection of mice to Fasciola hepatica

Rivera

Espino

2016

Experimental Parasitology

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Fasciola hepatica saposin-like protein-2 (FhSAP2) is a protein differentially expressed in various developmental stages of F. hepatica. Recombinant FhSAP2 has demonstrated the induction of partial protection in mice and rabbits when it is administered subcutaneously (SC) in Freund’s adjuvant. Because FhSAP2 is overexpressed in bacteria in the form of inclusion bodies (IBs), we isolated IBs expressing FhSAP2 and tested their immunogenicity when administered SC in mice emulsified in two different adjuvants: QS-21 and Montanide™ ISA720. Animals received three injections containing 20μg of protein two weeks apart and 4 weeks after the third injection, mice were infected with 10 F. hepatica metacercariae by oral route. The percentages of protection induced by FhSAP2-IBs were estimated to be between 60.0–62.5% when compared with adjuvant-vaccinated, infected controls. By determining the levels of IgG1 and IgG2a antibodies and IL-4 and IFNγ cytokines in the serum of experimental animals, it was found that both Th1 and Th2 immune responses were significantly increased in the FhSAP2-IBs vaccinated groups compared with the adjuvant-vaccinated, infected control groups. The adjuvant-vaccinated groups had significantly lower IgG1 to IgG2a ratios and lower IL-4 to IFNγ ratios than the FhSAP2-IBs vaccinated animals, which is indicative of higher levels of Th2 immune responses. Irrespective to the adjuvant used, animals vaccinated with FhSAP2-IBs exhibited significantly higher survival percentage and less liver damage than the adjuvant-control groups. This study suggests that FhSAP2 has potential as vaccine against F. hepatica and that the protection elicited by this molecule could be linked to a mechanism driven by the CD4-Th1 cells.

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“…Several different types of acidic/neutral cytosolic-, μ-, σ- and ω-class GSTs have been characterized in helminth parasites [17]. Most helminth GSTs are expressed in the parasite's parenchyme and subteguments, which suggests either that they interact with physiologically/pathologically active molecules during the phase II detoxification process, or that these enzymes are secreted and directly contact the host's defensive system [18]. Crystal structure and molecular modeling studies have revealed critical differences in their substrate and/or inhibitor specificity according to xenobiotic substrate-binding sites [19], [20], which implies that these isozymes might mediate different cellular homeostatic processes.…”

Section: Introductionmentioning

confidence: 99%

Differential Activation of Diverse Glutathione Transferases of Clonorchis sinensis in Response to the Host Bile and Oxidative Stressors

et al. 2013

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Background Clonorchis sinensis causes chronic cumulative infections in the human hepatobiliary tract and is intimately associated with cholangiocarcinoma. Approximately 35 million people are infected and 600 million people are at risk of infections worldwide. C. sinensis excretory-secretory products (ESP) constitute the first-line effector system affecting the host-parasite interrelationship by interacting with bile fluids and ductal epithelium. However, the secretory behavior of C. sinensis in an environment close to natural host conditions is unclear. C. sinensis differs from Fasciola hepatica in migration to, and maturation in, the hepatic bile duct, implying that protein profile of the ESP of these two trematodes might be different from each other.Methodology/Principal FindingsWe conducted systemic approaches to analyze the C. sinensis ESP proteome and the biological reactivity of C. sinensis glutathione transferases (GSTs), such as global expression patterns and induction profiles under oxidative stress and host bile. When we observed ex host excretion behavior of C. sinensis in the presence of 10% host bile, the global proteome pattern was not significantly altered, but the amount of secretory proteins was increased by approximately 3.5-fold. Bioactive molecules secreted by C. sinensis revealed universal/unique features in relation to its intraluminal hydrophobic residing niche. A total of 38 protein spots identified abundantly included enzymes involved in glucose metabolism (11 spots, 28.9%) and diverse-classes of glutathione transferases (GSTs; 10 spots, 26.3%). Cathepsin L/F (four spots, 10.5%) and transporter molecules (three spots, 7.9%) were also recognized. The universal secretory proteins found in other parasites, such as several enzymes involved in glucose metabolism and oxygen transporters, were commonly detected. C. sinensis secreted less cysteine proteases and fatty acid binding proteins compared to other tissue-invading or intravascular trematodes. Interestingly, secretion of a 28 kDa σ-class GST (Cs28σGST3) was significantly affected by the host bile, involving reduced secretion of the 28 kDa species and augmented secretion of Cs28σGST3-related high-molecular-weight 85 kDa protein. Oxidative stressors induced upregulated secretion of 28 kDa Cs28σGST3, but not an 85 kDa species. A secretory 26 kDa μ-class GST (Cs26μGST2) was increased upon treatment with oxidative stressors and bile juice, while another 28 kDa σ-class GST (Cs28σGST1) showed negligible responses.Conclusions/SignificanceOur results represent the first analysis of the genuine nature of the C. sinensis ESP proteome in the presence of host bile mimicking the natural host environments. The behavioral patterns of migration and maturation of C. sinensis in the bile ducts might contribute to the secretion of copious amounts of diverse GSTs, but a smaller quantity and fewer kinds of cysteine proteases. The Cs28σGST1 and its paralog(s) detoxify endogenous oxidative molecules, while Cs28σGST3 and Cs26μGST2 conjugate xenobiotics/hydrophobic...

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A recombinant thioredoxin-glutathione reductase from Fasciola hepatica induces a protective response in rabbits

Cited by 29 publications

References 37 publications

Identification of Thioredoxin Glutathione Reductase Inhibitors That Kill Cestode and Trematode Parasites

Identification of Thioredoxin Glutathione Reductase Inhibitors That Kill Cestode and Trematode Parasites

Adjuvant-enhanced antibody and cellular responses to inclusion bodies expressing FhSAP2 correlates with protection of mice to Fasciola hepatica

Differential Activation of Diverse Glutathione Transferases of Clonorchis sinensis in Response to the Host Bile and Oxidative Stressors

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