日本血吸虫童虫期别体被蛋白T辅助细胞表位预测及初步鉴定

发布时间：2018-05-06 09:21

  本文选题：日本血吸虫 + 体被分子　； 参考：《上海师范大学》2009年硕士论文

【摘要】： 日本血吸虫病是一种危害严重的人畜共患寄生虫病,严重影响我国包括农业和畜牧业在内的社会经济的可持续发展,应用抗血吸虫疫苗免疫人畜以阻断血吸虫病的传播是公认的长期有效的控制策略。安全高效实用的血吸虫分子疫苗的研发,由于鉴定保护性抗原遇到挑战,一直难以突破。以模拟辐照致弱尾蚴疫苗诱导产生的、由Th1细胞介导的、靶向童虫的高保护性免疫效应机制,来发现抗血吸虫的分子疫苗,可能是重要的突破口之一。本研究应用反向疫苗学技术从日本血吸虫转录本组中鉴别出童虫期别体表蛋白分子,预测其潜在的Th细胞表位,同时对其进行初步实验鉴定,为进一步发现源于血吸虫童虫体被分子的保护性T细胞抗原奠定基础。 本研究从国家人类基因组南方研究中心提供的日本血吸虫肝童虫期EST序列18,579条,从中筛选出童虫期别体表蛋白373条,对其进行ORF识别,用基于人工神经网络、隐马尔科夫模型、及权重矩阵分析上述序列的信号肽及跨膜结构,从373条日本血吸虫肝期童虫体被蛋白序列中筛得14条具有ORF结构及信号肽结构的序列。 应用免疫信息学技术进行候选序列的Th细胞表位预测。应用基于结合基序、矩阵法和人工神经网络等方法的多个服务器预测可与人源、鼠源MHCⅡ类分子结合的线性表位肽;应用基于分子动力学算法的受体配体嵌合空间结合模拟服务器,模拟肽与MHCⅡ分子的锚定情况,得到候选的Th细胞表位。结果预测得到了5个十五肽表位序列:NQFIRNLTLKIVLPE[源序列号AY813997;多萜基二磷酸寡聚糖蛋白葡萄糖基转移酶类似物];SSTSQDFPGVCQLCV[AY815893;原鞘脂激活蛋白];SSLVISYSTVDRLVC[源序列号AY815893;原鞘脂激活蛋白];SRNLTVLRTNVALRL[源序列号AY811606;秀丽隐杆线虫假象蛋白类似物]TVKFDKTVTCGGAYI[源序列号AAC00515;钙网织蛋白家族]。 用人工合成多肽法(四分臂偶联法合成到多聚赖氨酸骨架上)合成上述表位肽;MTT法改良技术CCK-8测定候选表位肽体外刺激免疫小鼠脾淋巴细胞增殖,结果初步鉴定获得了1个童虫期别蛋白候选Th细胞表位:SSLVISYSTVDRLVC[源序列号AY815893;原鞘脂激活蛋白]。 本研究采用免疫信息学理论预测技术结合体外试验检测技术,初步鉴定获得了日本血吸虫童虫期别体被蛋白分子Th细胞表位,为进一步筛选新的血吸虫童虫体被分子的保护性T细胞抗原奠定基础。
[Abstract]:Schistosomiasis japonicum is a serious zoonotic parasitic disease, which seriously affects the sustainable development of society and economy, including agriculture and animal husbandry. Anti-schistosomiasis vaccine is recognized as a long-term effective control strategy to prevent the transmission of schistosomiasis. The research and development of a safe, efficient and practical Schistosoma japonicum molecular vaccine has been difficult to break through due to the challenge of identification of protective antigen. The molecular vaccine against Schistosoma japonicum may be one of the important breakthroughs through the mechanism of high protective immunity induced by simulated irradiated attenuated cercariae vaccine and mediated by Th1 cells. In this study, reverse vaccinology was used to identify the allotropic protein molecules from Schistosoma japonicum transcripts, to predict the potential Th cell epitopes, and to identify the potential Th cell epitopes. It provides a basis for further discovery of protective T cell antigen derived from Schistosoma japonicum. In this study, 18579 EST sequences of Schistosoma japonicum liver stage were selected from 18579 EST sequences of Schistosoma japonicum liver from the National Center for the Southern Research of Human Genome, and 373 proteins were identified by ORF, using artificial neural network and Hidden Markov Model (Hidden Markov Model). The signal peptides and transmembrane structures of the above sequences were analyzed by weight matrix. Fourteen of 373 Schistosoma japonicum liver phase juvenile body protein sequences with ORF structure and signal peptide structure were obtained. Immunoinformatics was used to predict the Th cell epitopes of candidate sequences. Multiple servers based on binding motif, matrix method and artificial neural network were used to predict linear epitope peptides that could bind to human and mouse MHC class 鈪，

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