Lee Myoung-Ro, Kim Yu-Jung, Kim Dae-Won, Yoo Won Gi, Cho Shin-Hyeong, Hwang Kwang Yeon, Ju Jung-Won, Lee Won-Ja
Division of Malaria and Parasitic Diseases, Korea National Institute of Health, Osong, Republic of Korea.
Mol Biochem Parasitol. 2012 Mar-Apr;182(1-2):1-6. doi: 10.1016/j.molbiopara.2011.11.006. Epub 2011 Nov 15.
Clonorchis sinensis, the causative agent of clonorchiasis, is widespread in East and Southeast Asia, including China, Vietnam and the Republic of Korea. We identified antigenic proteins from adult C. sinensis liver flukes using immunoproteomic analysis. In this study, we found 23 candidate antigenic proteins with a pI in the range of 5.4-6.2 in total lysates of C. sinensis. The antigenic protein spots reacted against sera from clonorchiasis patients and were identified as cysteine proteases, glutathione transferases, gelsolin, propionyl-CoA carboxylase (PCC), prohibitin and 14-3-3 protein (14-3-3) using LC-coupled ESI-MS/MS and an EST database for C. sinensis. PCC and 14-3-3 were identified for the first time as serological antigens for the diagnosis of C. sinensis. To validate the antigenicity of PCC and 14-3-3, recombinant proteins were immunoblotted with sera from clonorchiasis patients. The structural, functional and immunological characteristics of the putative amino acid sequence were predicted by bioinformatics analysis. Our novel finding will contribute to the development of diagnostics for clonorchiasis. These results suggest that immunoproteomic approaches are valuable tools to identify antigens that could be used as targets for effective parasitic infection control strategies.
华支睾吸虫是华支睾吸虫病的病原体,广泛分布于东亚和东南亚,包括中国、越南和韩国。我们通过免疫蛋白质组学分析鉴定了成年华支睾吸虫肝吸虫的抗原蛋白。在本研究中,我们在华支睾吸虫的总裂解物中发现了23种候选抗原蛋白,其pI范围为5.4-6.2。这些抗原蛋白斑点与华支睾吸虫病患者的血清发生反应,并使用液相色谱-电喷雾串联质谱(LC-coupled ESI-MS/MS)和华支睾吸虫的EST数据库鉴定为半胱氨酸蛋白酶、谷胱甘肽转移酶、凝溶胶蛋白、丙酰辅酶A羧化酶(PCC)、抑制素和14-3-3蛋白(14-3-3)。PCC和14-3-3首次被鉴定为诊断华支睾吸虫的血清学抗原。为了验证PCC和14-3-3的抗原性,用华支睾吸虫病患者的血清对重组蛋白进行免疫印迹分析。通过生物信息学分析预测了假定氨基酸序列的结构、功能和免疫学特征。我们的新发现将有助于华支睾吸虫病诊断方法的开发。这些结果表明,免疫蛋白质组学方法是鉴定可作为有效控制寄生虫感染策略靶点的抗原的有价值工具。