Characterization of thioredoxin glutathione reductase in Schiotosoma japonicum

• Published in: Parasitology international Vol. 61; no. 3; pp. 475 - 480
• Main Authors: Han, Yanhui, Zhang, Min, Hong, Yang, Zhu, Zhu, Li, Dong, Li, Xiangrui, Fu, Zhiqiang, Lin, Jiaojiao
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Ireland Ltd 01.09.2012
• Subjects: adults; amino acids; Animals; antibodies; Clone; Cloning, Molecular; complementary DNA; disease control; DNA, Complementary - genetics; DNA, Helminth - genetics; eggs; enzyme activity; enzyme-linked immunosorbent assay; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Female; Gastroenterology and Hepatology; genes; Helminth Proteins - genetics; Helminth Proteins - metabolism; immunoglobulin G; Infectious Disease; interferon-gamma; Male; Mice; Mice, Inbred BALB C; Multienzyme Complexes - genetics; Multienzyme Complexes - metabolism; NADH, NADPH Oxidoreductases - genetics; NADH, NADPH Oxidoreductases - metabolism; open reading frames; parasitology; public health; quantitative polymerase chain reaction; Real-Time Polymerase Chain Reaction; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Schistosoma japonicum; Schistosoma japonicum - enzymology; Schistosoma japonicum - genetics; schistosomiasis; Schistosomiasis japonica - immunology; Schistosomiasis japonica - parasitology; Schistosomiasis japonica - pathology; screening; Sequence Analysis, DNA; Thioredoxin glutathione reductase; vaccination; Vaccination - methods; Vaccine; vaccines; China; Vaccine; Schistosoma japonicum; Thioredoxin glutathione reductase; Clone
• ISSN: 1383-5769; 1873-0329
• DOI: 10.1016/j.parint.2012.03.005

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 5.13 U/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.