Department of Parasitology and Immunology, School of Medicine, Yangzhou University, Yangzhou 225001, China.
Trans R Soc Trop Med Hyg. 2012 Jun;106(6):356-62. doi: 10.1016/j.trstmh.2012.02.008. Epub 2012 Apr 24.
The gene encoding surface antigen 2 (SAG2) or rhoptry protein 2 (ROP2) of Toxoplasma gondii was cloned into the plasmid pGEX-4T-1 and subsequently expressed in Escherichia coli as a glutathione-s-transferase (GST) fusion protein. The characteristics of purified GST-SAG2 or GST-ROP2 were analysed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analysis. The specific IgG of a panel of serum samples provided by the National Institute for the Control of Pharmaceutical and Biological Products were tested with commercial ELISA and the lateral flow immunoassay (LFIA) based on GST-SAG2, GST-ROP2 or GST-SAG2+ROP2. A total of 1096 sera and saliva samples from pregnant women were tested by GST-SAG2+ROP2-LFIA. In total, 20 T. gondii IgM positive sera (1.82%), 81 T. gondii IgG positive sera (7.4%) and 23 T. gondii IgA positive saliva (2.1%) were finally confirmed. The SAG2+ROP2 specific IgG and IFN-γ producing CD8+ T cells were induced in mice immunised with GST-SAG2+ROP2. The results indicate that GST-SAG2+ROP2 protein can be used as an antigen for diagnosing T. gondii infection and provide a strategy for development of subunit vaccines for protection against T. gondii infection.
弓形虫表面抗原 2（SAG2）或速殖子蛋白 2（ROP2）基因被克隆到质粒 pGEX-4T-1 中，并在大肠杆菌中作为谷胱甘肽-S-转移酶（GST）融合蛋白表达。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）和免疫印迹分析对纯化的 GST-SAG2 或 GST-ROP2 的特性进行了分析。使用商业 ELISA 和基于 GST-SAG2、GST-ROP2 或 GST-SAG2+ROP2 的侧向流动免疫分析（LFIA）对国家药品和生物制品控制研究所提供的一组血清样本中的特异性 IgG 进行了检测。对 1096 份孕妇血清和唾液样本进行了 GST-SAG2+ROP2-LFIA 检测。最终共确认了 20 份 T. gondii IgM 阳性血清（1.82%）、81 份 T. gondii IgG 阳性血清（7.4%）和 23 份 T. gondii IgA 阳性唾液（2.1%）。用 GST-SAG2+ROP2 免疫的小鼠中诱导了 SAG2+ROP2 特异性 IgG 和 IFN-γ 产生的 CD8+T 细胞。结果表明，GST-SAG2+ROP2 蛋白可用作诊断 T. gondii 感染的抗原，并为开发针对 T. gondii 感染的亚单位疫苗提供了策略。
