【摘要】 Label-free定性技术是基于质谱分析蛋白质组学的研究方法,适用于样品量小或样品稀缺物种的蛋白质组学研究。为了筛选猪囊尾蚴病的特异性抗原诊断标识,首先利用高效液相色谱-串联质谱系统对虫体和患者血清进行全蛋白定性分析,然后筛选囊虫的外分泌蛋白。结果表明,人感染猪囊虫后,囊虫的外分泌蛋白刺激宿主机体产生免疫应答,参与寄生虫逃避宿主的免疫机制。为探究囊虫外分泌蛋白在免疫逃避机制中的意义,通过基因本体论(GO)、直系同源簇(COG)、京都基因与基因组百科全书(KEGG)富集等分析猪源囊虫和患者血清蛋白质组学,富集到参与细胞内进程、单体进程、生物学调节等蛋白,主要参与补体与凝血级联、ECM受体相互作用等信号通路。通过预测患者血清的蛋白质结构域(Domain),发现蛋白A0A0R3X5M7是囊虫外分泌蛋白。本试验阐明了囊虫外分泌蛋白对宿主机体的影响,为研究囊虫外分泌蛋白提供基础,为猪囊尾蚴病不同感染时期诊断标识的筛选提供思路。更多还原

【Abstract】 Label-free qualitative technique is a proteomic research method based on mass spectrometry, which is suitable for proteomic study of species with small or scarce samples. To screen the specific antigen diagnostic marker of porcine Cysticercosis firstly, the whole protein of the worm and the patient’s serum samples were qualitatively analyzed by high performance liquid chromatography-tandem mass spectrometry system, and then the exocrine protein of Cysticercus was screened. The results showed that the exocrine protein of Cysticercus stimulated the host immune response and participated in the immune mechanism of parasite escaping after human infection with Cysticercus. The methods of gene ontology(GO), clusters of orthologous groups of proteins(COG), kyoto encyclopedia of gene and genome(KEGG) enrichment were used to analyze the serum proteomics of Cysticercus cellulosae and patients. The enriched proteins participated in the intracellular process, monomer process, biological regulation and other biological processes, mainly involved in complement and coagulation cascade, ECM receptor interaction and other signal pathways, which is to explore the significance of cysticercus exocrine proteins in immune escape mechanism. By predicting the protein domain(Domain) of the patient’s serum, we found that the protein A0 A0 R3 X5 M7 was an exocrine protein of Cysticercus. This study clarifies the effect of Cysticercus exocrine protein on host, and will provide the basis for the study of Cysticercus exocrine protein and ideas for the screening of diagnostic markers of Cysticercooids cellulosae in different infection stages.更多还原