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Identification and immunological characterization of thioredoxin transmembrane-related protein from Clonorchis sinensis.
- Source :
-
Parasitology research [Parasitol Res] 2013 Apr; Vol. 112 (4), pp. 1729-36. Date of Electronic Publication: 2013 Feb 13. - Publication Year :
- 2013
-
Abstract
- Thioredoxin transmembrane related protein (TMX), a member of thioredoxin superfamily, is localized to the endoplasmic reticulum and possesses a thioredoxin-like domain that plays an important role as an oxidoreductase. The functions of TMX in Clonorchis sinensis remain to be elucidated. In this study, we cloned and characterized a novel TMX of C. sinensis (CsTMX). The CsTMX cDNA sequence contained a 414-nucleotide open-reading frame encoding a protein of 137 amino acids. A thioredoxin domain was found in the position of aa21-117 and contained the putative active-site motif Cys-Pro-Ala-Cys. BLASTx analysis showed that CsTMX shared 39-57% amino acid identities with TMX of other organisms. Quantitative RT-PCR analysis demonstrated that CsTMX was differentially transcribed, with the highest level of expression in the adult worm stage and the lowest expression in egg stage. In addition, immunofluorescence assay showed CsTMX was localized in the tegument, vitelline gland, intestine, and intrauterine eggs of adult worm. Besides, immunoblot assay revealed that the recombinant CsTMX (rCsTMX) could be recognized by the sera from rats infected with C. sinensis and the sera from rats immunized by excretory-secretory products. Furthermore, analysis of the antibody isotype profile revealed that rats subcutaneously immunized with rCsTMX developed rCsTMX-specific antibody, which is dominance of IgG2a in sera. Meanwhile, production of IFN-γ was elevated strongly in the supernatants of spleen cell. The results collectively indicated that CsTMX might play an important role in the host-parasite interaction, as well as CsTMX probably involved in immunoregulation of host by inducing Th1-type dominated immune response in rats.
- Subjects :
- Amino Acid Sequence
Animal Structures chemistry
Animals
Antibodies, Helminth blood
Catalytic Domain
Cloning, Molecular
Clonorchiasis immunology
Clonorchiasis parasitology
Clonorchis sinensis immunology
DNA, Complementary
Disease Models, Animal
Fluorescent Antibody Technique
Gene Expression Profiling
Immunoglobulin G blood
Membrane Proteins immunology
Molecular Sequence Data
Open Reading Frames
Protein Structure, Tertiary
Rats
Rats, Wistar
Real-Time Polymerase Chain Reaction
Sequence Homology, Amino Acid
Thioredoxins immunology
Clonorchis sinensis chemistry
Clonorchis sinensis genetics
Membrane Proteins analysis
Membrane Proteins genetics
Thioredoxins analysis
Thioredoxins genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1432-1955
- Volume :
- 112
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Parasitology research
- Publication Type :
- Academic Journal
- Accession number :
- 23403994
- Full Text :
- https://doi.org/10.1007/s00436-013-3331-5