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Molecular characterization of thyroid hormone receptor beta from Schistosoma japonicum and assessment of its potential as a vaccine candidate antigen against schistosomiasis in BALB/c mice
Qiu, Chunhui ; Liu, Shengfa ; Liu SF(刘生发) ; Hong, Yang ; Fu, Zhiqiang ; Wei, Meimei ; Ai, Dezhou ; Lin, Jiaojiao
2012-08-13
关键词NUCLEAR RECEPTOR GENE-EXPRESSION MANSONI IDENTIFICATION TUBERCULOSIS CYTOKINES INVITRO IMMUNE
英文摘要Background: Thyroid hormones (TH) modulate growth, development and differentiation and metabolic processes by interacting with thyroid hormone receptors (THRs). The purpose of this study was to identify a novel thyroid hormone receptor beta encoding gene of Schistosoma japonicum (SjTHR beta) and to investigate its potential as a vaccine candidate antigen against schistosomiasis in BALB/c mice. Methods: The full-length cDNA sequence of SjTHR beta, its gene organization, and its transcript levels were characterized, and the phylogenetic relationship between THR, RAR and RXR from other organisms were analysis, the ability of this protein binding to a conserved DNA core motif, and its potential as a vaccine candidate antigen against schistosomiasis in BALB/c mice were evaluated. Results: The SjTHR beta cDNA was cloned, verified by 5' and 3' Rapid Amplification of cDNA Ends and shown to be polyadenylated at the 3' end, suggesting the transcript is full-length. SjTHR beta is homologous to THRs from other species and has a predicted conservative DNA binding domain and ligand binding domain that normally characterizes these receptors. A comparative quantitative PCR analysis showed that SjTHR beta was the highest expressed in 21d worms and the lowest in 7 d and 13 d schistosomula. The cDNA corresponding to DNA binding domain (SjTHR beta-DBD) and ligand binding domain (SjTHR beta-LBD) were cloned and subsequently expressed in E coli. The expressed proteins were used to immunize mice and generate specific serum against recombinant SjTHR beta (rSjTHR beta). Western blotting revealed that anti-rSjTHR beta-LBD serum recognized two protein bands in extracts from 21 d worm with molecular sizes of approximately 95 kDa and 72 kDa. Electrophoretic mobility shift assay (EMSA) analysis showed that rSjTHR beta-DBD could bind to a conserved DNA core motif. Immunization of BALB/c mice with rSjTHR beta-LBD could induce partial protective efficacy(27.52% worm reduction and 29.50% liver eggs reduction) against schistosome infection. Enzyme-linked immunosorbent assay showed that mice vaccinated with recombinant SjTHR beta-LBD (rSjTHR beta-LBD) generated increased levels of specific IgG, IgG1 and IgG2a antibody. Bio-plex analysis demonstrated that rSjTHR beta-LBD induced considerably higher levels of T helper 1 cytokines (IL-2, IL-12 and TNF-alpha) than T helper 2 cytokines (IL-10, IL-4), suggesting that rSjTHR beta-LBD vaccination could stimulate mixed Th1/Th2 types with Th1 dominant immune responses. Conclusions: Our study presented here identified SjTHR beta as a new schistosome THR that might play an important role in host-parasite interaction and be a vaccine candidate for schistosomiasis.; National Natural Science Foundation of China [31172315]; Special Fund for Agro-Scientific Research in the Public Interest [200903036]
语种英语
出版者BIOMED CENTRAL LTD
内容类型期刊论文
源URL[http://dx.doi.org/10.1186/1756-3305-5-172]  
专题生命科学-已发表论文
推荐引用方式
GB/T 7714
Qiu, Chunhui,Liu, Shengfa,Liu SF,et al. Molecular characterization of thyroid hormone receptor beta from Schistosoma japonicum and assessment of its potential as a vaccine candidate antigen against schistosomiasis in BALB/c mice[J],2012.
APA Qiu, Chunhui.,Liu, Shengfa.,刘生发.,Hong, Yang.,Fu, Zhiqiang.,...&Lin, Jiaojiao.(2012).Molecular characterization of thyroid hormone receptor beta from Schistosoma japonicum and assessment of its potential as a vaccine candidate antigen against schistosomiasis in BALB/c mice..
MLA Qiu, Chunhui,et al."Molecular characterization of thyroid hormone receptor beta from Schistosoma japonicum and assessment of its potential as a vaccine candidate antigen against schistosomiasis in BALB/c mice".(2012).
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