title: Characterization of thioredoxin glutathione reductase in Schiotosoma japonicum.
abstract: Schistosomiasis is one of the most prevalent and serious parasitic diseases in the world and remains an important public health problem in China. Screening and discovery of an effective vaccine candidate or new drug target is crucial for the control of this disease. In this study, we cloned a cDNA encoding Schistosoma japonicum (S. japonicum) thioredoxin glutathione reductase (SjTGR) from the cDNA of 42-day-old adult worms. The open reading frame (ORF) of the gene was 1791 base pairs (bp) encoding a protein of 596 amino acids. SjTGR was subcloned into pET-32a (+) and expressed in Escherichia coli (E. coli) BL21 (DE3). The recombinant protein rSjTGR exhibited enzymatic activity of 5.13U/mg with DTNB as the substrate, and showed strong immunogenecity. Real-time PCR results indicated that SjTGR was expressed at a higher level in 35-day-old schistosome worms in transcript. We vaccinated BALB/c mice with rSjTGR in combination with MONTANIDE™ ISA 206 VG (ISA 206) and observed a 33.50% to 36.51% (P<0.01) decrease in the adult worm burden and a 33.73%to 43.44% (P<0.01) decrease in the number of eggs counted compared to the ISA 206 or blank control groups in two independent vaccination tests. ELISA analysis demonstrated that rSjTGR induced a high level of SjTGR-specific IgG, IgG1, and IgG 2a antibodies and induced elevated production of IFN-γ. This study provides the basis for further investigations into the biological function of SjTGR and further evaluation of the potential use of this molecule as a vaccine candidate or new drug target is warranted.
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai 200241, PR China.
authors.forename: Yanhui
authors.initials: Y
authors.lastname: Han
authors.forename: Min
authors.initials: M
authors.lastname: Zhang
authors.forename: Yang
authors.initials: Y
authors.lastname: Hong
authors.forename: Zhu
authors.initials: Z
authors.lastname: Zhu
authors.forename: Dong
authors.initials: D
authors.lastname: Li
authors.forename: Xiangrui
authors.initials: X
authors.lastname: Li
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Fu
authors.forename: Jiaojiao
authors.initials: J
authors.lastname: Lin
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D018509:DNA, Helminth / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D015801:Helminth Proteins / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009097:Multienzyme Complexes / Q000235:genetics / Q000378:metabolism*; D009247:NADH, NADPH Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012549:Schistosoma japonicum / Q000201:enzymology* / Q000235:genetics; D012554:Schistosomiasis japonica / Q000276:immunology / Q000469:parasitology / Q000473:pathology; D017422:Sequence Analysis, DNA; D014611:Vaccination / Q000379:methods
chemical_list: D018076:DNA, Complementary; D018509:DNA, Helminth; D015801:Helminth Proteins; D009097:Multienzyme Complexes; D011994:Recombinant Proteins; D009247:NADH, NADPH Oxidoreductases; C466433:thioredoxin glutathione reductase