Characterization of the secreted cathepsin B cysteine proteases family of the carcinogenic liver fluke Clonorchis sinensis
Characterization of the secreted cathepsin B cysteine proteases family of the carcinogenic liver fluke Clonorchis sinensis
复制标题
致癌性肝吸虫华支睾吸虫分泌型组织蛋白酶 B 半胱氨酸蛋白酶家族的表征
DOI：
10.1007/s00436-014-4006-6
复制
发表时间：
2014-09-01
影响因子：
2
通讯作者：
Yu, Xinbing
中科院分区：
文献类型：
--
作者：
Chen, Wenjun;Wang, Xiaoyun;Yu, Xinbing
中华绒螯蟹排泄分泌产物（ESP）因其作为疫苗候选物和药物靶点的潜力而备受关注。防治冬虫夏草在这项研究中，我们广泛地描绘了四个C。中国人组织蛋白酶B半胱氨酸蛋白酶（CsCB 1、CsCB 2、CsCB 3和CsCB 4）。生物信息学分析表明，所有CsCB的N端均含有信号肽。在CsCB序列中发现了功能结构域和残基。我们表达了四种CsCB，并分析了疫苗试验后的免疫反应。重组CsCB可诱导高滴度的IgG，表明CsCB家族具有较高的免疫原性。ELISA结果显示，免疫后小鼠IgG 1和IgG 2a水平均明显升高，提示CsCB家族可诱导Th 1/Th 2联合免疫应答。RT-PCR和Western blotting结果均证实了4种CsCB在C.中国生活史更重要的是，我们验证了我们的假设，CsCB是C。中华绒螯蟹排泄/分泌产物。CsCB可被C.中华绒螯蟹感染的血清中，表明分泌的CsCB是C.中华感染通过比较CsCB组与对照组的虫载量和每克虫卵数（EPG）来评价CsCB的保护效果，CsCB 2和CsCB 3组的虫载量（P <0.01）和EPG（P < 0.01）均显著低于对照组。总之，我们首次对分泌型组织蛋白酶B半胱氨酸蛋白酶家族进行了谱分析，并证明所有CsCB家族均为C。可能调节宿主免疫反应的中华绒螯蟹排泄/分泌产物。
Clonorchis sinensis excretory/secretory products (ESP) have gained high attentions because of their potential to be vaccine candidates and drug targets in C. sinensis prevention. In this study, we extensively profiled the characteristics of four C. sinensis cathepsin B cysteine proteases (CsCB1, CsCB2, CsCB3, and CsCB4). Bioinformatics analysis showed all CsCBs contained signal peptides at the N-terminal. Functional domains and residues were found in CsCB sequences. We expressed four CsCBs and profiled immune responses followed by vaccine trials. Recombinant CsCBs could induce high IgG titers, indicating high immunogenicity of CsCB family. Additionally, ELISA results showed that both IgG1 and IgG2a levels apparently increased post-immunization with all four CsCBs, showing that combined Th1/Th2 immune responses were triggered by CsCB family. Both Real-time polymerase chain reaction (RT-PCR) and Western blotting confirmed that four CsCBs have distinct expression patterns in C. sinensis life stages. More importantly, we validated our hypothesis that CsCBs were C. sinensis excretory/secretory products. CsCBs could be recognized by C. sinensis-infected sera throughout the infection period, indicating that secreted CsCBs are immune triggers during C. sinensis infection. The protective effect was assessed by comparing the worm burden and egg per gram (EPG) between CsCB group and control group, showing that worm burden (P < 0.01) and EPG (P < 0.01) in CsCB2 and CsCB3 groups were significantly lower than in control group. In conclusion, we profiled secreted cathepsin B cysteine proteases family for the first time and demonstrated that all CsCB family were C. sinensis excretory/secretory products that may regulate host immune responses.