National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.
Parasitol Res. 2011 Sep;109(3):899-911. doi: 10.1007/s00436-011-2332-5. Epub 2011 Apr 2.
The characterization of the cross-reactive and species-specific antigens of Neospora caninum and Toxoplasma gondii is important in the exploration to determine the common mechanisms of parasite-host interaction and to improve the serological diagnosis; it is also useful for the selection of the cross-reactive antigens that could be used in the development of vaccines or drugs for controlling the diseases caused by these two parasites. In this study, cross-reactive and species-specific antigens between N. caninum and T. gondii tachyzoites were comprehensively investigated using a proteomics approach with the application of two-dimensional gel electrophoresis, immunoblot analysis, matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS), and MALDI-TOF/TOF-MS analysis. Immunoblotting and mass spectrometry analysis revealed that at least 42 individual protein spots of N. caninum were reacted with the anti-N. caninum serum, among which at least 18 protein spots were cross-reacted with the anti-T. gondii serum. Moreover, at least 31 protein spots of T. gondii were reacted with the anti-T. gondii serum, among which at least 19 protein spots were cross-reacted with the anti-N. caninum serum. Furthermore, some new specific proteins were also identified in the N. caninum protein profile by searching Toxoplasma sequences or sequences from other organisms. This study substantiates the usefulness of proteomics in the immunoscreening of the cross-reactive or species-specific antigens of both parasites. In addition, the present study showed that there was significant homology in the antigenic proteome profiles between the two parasites. These observations have implications for the design of multicomponent common vaccines against both parasite infections.
刚地弓形虫和新孢子虫的交叉反应性和种特异性抗原的特性研究对于探索寄生虫与宿主相互作用的共同机制和改进血清学诊断非常重要；它还有助于选择交叉反应性抗原，这些抗原可用于开发控制这两种寄生虫引起的疾病的疫苗或药物。在这项研究中，应用二维凝胶电泳、免疫印迹分析、基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF-MS）和 MALDI-TOF/TOF-MS 分析等蛋白质组学方法，全面研究了刚地弓形虫和新孢子虫速殖子之间的交叉反应性和种特异性抗原。免疫印迹和质谱分析表明，至少有 42 个新孢子虫的个体蛋白斑点与抗新孢子虫血清反应，其中至少有 18 个蛋白斑点与抗刚地弓形虫血清反应。此外，至少有 31 个刚地弓形虫的蛋白斑点与抗刚地弓形虫血清反应，其中至少有 19 个蛋白斑点与抗新孢子虫血清反应。此外，通过搜索弓形虫序列或其他生物体的序列，还在新孢子虫蛋白图谱中鉴定出一些新的特异性蛋白。本研究证实了蛋白质组学在免疫筛选两种寄生虫的交叉反应性或种特异性抗原中的有用性。此外，本研究表明，两种寄生虫的抗原蛋白图谱之间存在显著的同源性。这些观察结果对设计针对两种寄生虫感染的多成分通用疫苗具有重要意义。
