血吸虫成虫排泄分泌抗原的组成及免疫反应性分析

Analysis of the proteomics and immunoreactivities of excretory-secretory antigens of Schistosoma japonicum adult worms

目的分析日本血吸虫成虫排泄分泌抗原的组成及其与血吸虫感染兔疗前、疗后配对血清的免疫反应特征。方法采用7cm(pH5～8)的IPG和12%SDS-PAGE对血吸虫成虫排泄分泌抗原的组成进行双向凝胶电泳分析,并用免疫印迹试验检测排泄分泌抗原各蛋白点与疗前、疗后血清的反应性;采用PDQuest8.0图像分析软件对二维图像斑点进行分析,寻找差异反应蛋白点。结果血吸虫成虫排泄分泌抗原双向凝胶电泳图谱主要由215个蛋白斑点组成,其中16个斑点大、染色深,可能为高丰度的循环抗原;免疫印迹显示可被感染6周兔血清识别,但不能被治疗12周兔血清识别的蛋白斑点有84个,与感染6周兔血清的反应强度比与治疗12周兔血清的反应强度高2倍的蛋白点有38个。结论血吸虫成虫排泄分泌抗原中存在高丰度的能被短程抗体识别的抗原分子。

Objective To analyze the components of excretory-secretory antigens of Schistosoma japonicum adult worms, and characterize the immune reaction in matching sera of rabbits infected with S. japonicum before and after treatment. Methods The excretory secretory antigens of S. japonicum adult worms were analyzed by two dimensional gel electrophoresis （2 DE） with 7-cm immobilized pH gradient （IPG） gels （pHS-8） and 12% SDS-PAGE. Protein spots on SDS-PAGE gels were transferred to a nitrocellulose membrane by electro-blotting and recognized with matching sera of rabbits infected with S. japonicum before and after treatment. Differences in spots were analyzed with PDQuest8. 0 image analysis software. Results A total of 215 protein spots was found on the 2 DE gel of excretory secretory antigens of adult worms; of these spots, 16 were larger and stained darker, which may indicate an abundance circulating antigens of S. japonicum. The results of Western blotting showed that 84 protein spots were recognized by the serum of rabbits 6 weeks after infection but were not recognized by the serum 12 weeks after treatment. For 38 spots, the intensity of the response to the serum of rabbits 6 weeks after infection was twice that of the response to serum of the same rabbits 12 weeks after treatment. Conclusion There were abundant circulating antigens in the excretory-secretory antigens of S. japonicum adult worms, and some antigens may induce short term antibodies in patients with schistosomiasis.