Key Laboratory of Zoonoses of Agriculture Ministry, Lanzhou Veterinary Research Institute, CAAS, Lanzhou, Gansu, China.
Parasitol Res. 2010 Apr;106(5):1151-7. doi: 10.1007/s00436-010-1786-1. Epub 2010 Feb 23.
Taenia solium is a cestode parasite that causes cysticercosis in humans and pigs. TSOL18 has been identified as a host-protective oncosphere antigen. To obtain mouse monoclonal antibodies (mAbs) against TSOL18 and to map its antigenic epitopes are potentials to develop a vaccine for the prevention of T. solium infection. In this study, mAbs were produced by the hybridoma technique using purified glycosylated TSOL18 produced in Pichia pastoris as the immunogen. mAb was used to define the B-cell epitopes of TSOL18 with phage-displayed random dodecapeptide library (Ph.D.-12), and some of the positive phage clones were sequenced and analyzed. The predominant mimotopes were ETTKLQRFQAML (L1) found in 83%, followed by DHTXF in 15% (L2: DHTLFAASHNHR, DHTLFSTGHSHG, and DHTFMQRYHTHQ). Comparison of the peptide sequences with native TSOL18 protein sequence using Clustal W software showed that they did not completely match, suggesting that the ETTKLQRFQAML and DHTXF sequences should be conformational epitopes. The sera of mice immunized with the selected phage clones obviously recognized the TSOL18 protein. Meanwhile, sera collected from TSOL18-vaccinated pigs reacted to both epitopes in enzyme-linked-immunosorbent serologic assay test. Our work demonstrated that the antigenic epitope could be mapped through screening the phage-displayed peptide libraries with mAbs and a mimotope of TSOL18, which could provide an alternative approach for the diagnosis and development of a vaccine for T. solium.
猪带绦虫是一种引起人体和猪囊尾蚴病的绦虫寄生虫。TSOL18 已被确定为一种宿主保护性的原头蚴抗原。获得针对 TSOL18 的小鼠单克隆抗体 (mAbs) 并绘制其抗原表位图是开发预防猪带绦虫感染疫苗的潜力。在这项研究中,使用毕赤酵母表达的纯化糖基化 TSOL18 作为免疫原,通过杂交瘤技术产生 mAb。mAb 用于使用噬菌体展示随机十二肽文库 (Ph.D.-12) 定义 TSOL18 的 B 细胞表位,并对一些阳性噬菌体克隆进行测序和分析。主要的模拟表位是在 83%的克隆中发现的 ETTKLQRFQAML (L1),其次是 DHTXF (L2: DHTLFAASHNHR、DHTLFSTGHSHG 和 DHTFMQRYHTHQ)。使用 Clustal W 软件将肽序列与天然 TSOL18 蛋白序列进行比较表明,它们不完全匹配,这表明 ETTKLQRFQAML 和 DHTXF 序列应该是构象表位。用选定的噬菌体克隆免疫的小鼠血清明显识别 TSOL18 蛋白。同时,从 TSOL18 疫苗接种的猪采集的血清在酶联免疫吸附血清学检测试验中对两个表位均有反应。我们的工作表明,通过用 mAbs 和 TSOL18 的模拟表位筛选噬菌体展示肽文库,可以绘制抗原表位,为猪带绦虫的诊断和疫苗开发提供一种替代方法。