title: Identification and immunological characterization of thioredoxin transmembrane-related protein from Clonorchis sinensis.
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.
authors: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, 510080, People's Republic of China.
authors: Chenhui
authors: C
authors: Zhou
authors: Meng
authors: M
authors: Bian
authors: Hua
authors: H
authors: Liao
authors: Qiong
authors: Q
authors: Mao
authors: Ran
authors: R
authors: Li
authors: Juanjuan
authors: J
authors: Zhou
authors: Xiaoyun
authors: X
authors: Wang
authors: Shan
authors: S
authors: Li
authors: Chi
authors: C
authors: Liang
authors: Xuerong
authors: X
authors: Li
authors: Yan
authors: Y
authors: Huang
authors: Xinbing
authors: X
authors: Yu
mesh_terms: D000595:Amino Acid Sequence; D000825:Animal Structures / Q000737:chemistry; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D020134:Catalytic Domain; D003001:Cloning, Molecular; D003003:Clonorchiasis / Q000276:immunology / Q000469:parasitology; D003004:Clonorchis sinensis / Q000737:chemistry* / Q000235:genetics* / Q000276:immunology; D018076:DNA, Complementary; D004195:Disease Models, Animal; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D007074:Immunoglobulin G / Q000097:blood; D008565:Membrane Proteins / Q000032:analysis* / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D017434:Protein Structure, Tertiary; D051381:Rats; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013879:Thioredoxins / Q000032:analysis* / Q000235:genetics* / Q000276:immunology
chemical_list: D000909:Antibodies, Helminth; D018076:DNA, Complementary; D007074:Immunoglobulin G; D008565:Membrane Proteins; D013879:Thioredoxins
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