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.
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.
复制标题
日本血吸虫甲状腺激素受体β的分子特征及其作为 BALB/c 小鼠血吸虫病疫苗候选抗原的潜力
DOI：
10.1186/1756-3305-5-172
复制
发表时间：
2012-08-13
影响因子：
3.2
通讯作者：
Lin J
中科院分区：
文献类型：
--
作者：
Qiu C;Liu S;Hong Y;Fu Z;Wei M;Ai D;Lin J
背景
甲状腺激素 (TH) 通过与甲状腺激素受体 (THR) 相互作用来调节生长、发育、分化和代谢过程。本研究的目的是鉴定日本血吸虫的新型甲状腺激素受体β编码基因（SjTHRβ），并研究其作为 BALB/c 小鼠血吸虫病疫苗候选抗原的潜力。
方法
对SjTHRβ的全长cDNA序列、其基因结构和转录水平进行了表征，并分析了来自其他生物体的THR、RAR和RXR之间的系统发育关系，评估了该蛋白与保守DNA核心基序结合的能力，及其作为BALB/c小鼠血吸虫病疫苗候选抗原的潜力。
结果
SjTHRβ cDNA 被克隆，通过 cDNA 末端的 5' 和 3' 快速扩增进行验证，并显示在 3' 端被聚腺苷酸化，表明转录物是全长的。 SjTHRβ 与其他物种的 THR 同源，并具有预测的保守 DNA 结合域和配体结合域，通常表征这些受体。比较定量PCR分析表明，SjTHRβ在21d的血吸虫中表达量最高，在7d和13d的血吸虫中表达最低。克隆对应于 DNA 结合结构域 (SjTHRβ-DBD) 和配体结合结构域 (SjTHRβ-LBD) 的 cDNA，并随后在大肠杆菌中表达。表达的蛋白用于免疫小鼠并产生针对重组 SjTHRβ (rSjTHRβ) 的特异性血清。蛋白质印迹显示，抗 rSjTHRβ-LBD 血清可识别 21 d 蠕虫提取物中的两条蛋白条带，分子大小约为 95 kDa 和 72 kDa。电泳迁移率变动分析 (EMSA) 分析表明 rSjTHRβ-DBD 可以结合保守的 DNA 核心基序。 rSjTHRβ-LBD 免疫 BALB/c 小鼠可产生针对血吸虫感染的部分保护效果（蠕虫减少 27.52%，肝卵减少 29.50%）。酶联免疫吸附测定显示，接种重组 SjTHRβ-LBD (rSjTHRβ-LBD) 的小鼠产生了更高水平的特异性 IgG、IgG1 和 IgG2a 抗体。 Bio-plex 分析表明，rSjTHRβ-LBD 诱导的 T 辅助细胞 1 细胞因子（IL-2、IL-12 和 TNF-α）水平明显高于 T 辅助细胞 2 细胞因子（IL-10、IL-4），表明 rSjTHRβ-LBD 疫苗接种可以刺激混合 Th1/Th2 类型和 Th1 主导免疫反应。
结论
我们在此提出的研究确定了 SjTHRβ 是一种新的血吸虫 THR，它可能在宿主-寄生虫相互作用中发挥重要作用，并成为血吸虫病的候选疫苗。
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 (SjTHRβ) and to investigate its potential as a vaccine candidate antigen against schistosomiasis in BALB/c mice.
Methods
The full-length cDNA sequence of SjTHRβ, 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 SjTHRβ 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. SjTHRβ 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 SjTHRβ was the highest expressed in 21d worms and the lowest in 7 d and 13 d schistosomula. The cDNA corresponding to DNA binding domain (SjTHRβ-DBD) and ligand binding domain (SjTHRβ-LBD) were cloned and subsequently expressed in E coli. The expressed proteins were used to immunize mice and generate specific serum against recombinant SjTHRβ (rSjTHRβ). Western blotting revealed that anti-rSjTHRβ-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 rSjTHRβ-DBD could bind to a conserved DNA core motif. Immunization of BALB/c mice with rSjTHRβ-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 SjTHRβ-LBD (rSjTHRβ-LBD) generated increased levels of specific IgG, IgG1 and IgG2a antibody. Bio-plex analysis demonstrated that rSjTHRβ-LBD induced considerably higher levels of T helper 1 cytokines (IL-2, IL-12 and TNF-α) than T helper 2 cytokines (IL-10, IL-4), suggesting that rSjTHRβ-LBD vaccination could stimulate mixed Th1/Th2 types with Th1 dominant immune responses.
Conclusions
Our study presented here identified SjTHRβ as a new schistosome THR that might play an important role in host-parasite interaction and be a vaccine candidate for schistosomiasis.
登录
查看更多内容
影响因子：
4.4
作者：
Gobert GN;Moertel L;Brindley PJ;McManus DP
通讯作者：
McManus DP
影响因子：
56.9
作者：
MORGAN, DA;RUSCETTI, FW;GALLO, R
通讯作者：
GALLO, R
影响因子：
2.7
作者：
van der Werf, MJ;de Vlas, SJ;Engels, D
通讯作者：
Engels, D
DOI：
10.1073/pnas.0904064106
发表时间：
2009-06-09
影响因子：
11.1
作者：
Wang, Zhu;Zhou, X. Edward;Mangelsdorf, David J.
通讯作者：
Mangelsdorf, David J.
影响因子：
5.4
作者：
Wu, Wenjie;Niles, Edward G.;LoVerde, Philip T.
通讯作者：
LoVerde, Philip T.