本文選題：日本血吸蟲 + 體被分子��； 參考：《上海師范大學(xué)》2009年碩士論文

【摘要】： 日本血吸蟲病是一種危害嚴(yán)重的人畜共患寄生蟲病,嚴(yán)重影響我國(guó)包括農(nóng)業(yè)和畜牧業(yè)在內(nèi)的社會(huì)經(jīng)濟(jì)的可持續(xù)發(fā)展,應(yīng)用抗血吸蟲疫苗免疫人畜以阻斷血吸蟲病的傳播是公認(rèn)的長(zhǎng)期有效的控制策略。安全高效實(shí)用的血吸蟲分子疫苗的研發(fā),由于鑒定保護(hù)性抗原遇到挑戰(zhàn),一直難以突破。以模擬輻照致弱尾蚴疫苗誘導(dǎo)產(chǎn)生的、由Th1細(xì)胞介導(dǎo)的、靶向童蟲的高保護(hù)性免疫效應(yīng)機(jī)制,來發(fā)現(xiàn)抗血吸蟲的分子疫苗,可能是重要的突破口之一。本研究應(yīng)用反向疫苗學(xué)技術(shù)從日本血吸蟲轉(zhuǎn)錄本組中鑒別出童蟲期別體表蛋白分子,預(yù)測(cè)其潛在的Th細(xì)胞表位,同時(shí)對(duì)其進(jìn)行初步實(shí)驗(yàn)鑒定,為進(jìn)一步發(fā)現(xiàn)源于血吸蟲童蟲體被分子的保護(hù)性T細(xì)胞抗原奠定基礎(chǔ)。 本研究從國(guó)家人類基因組南方研究中心提供的日本血吸蟲肝童蟲期EST序列18,579條,從中篩選出童蟲期別體表蛋白373條,對(duì)其進(jìn)行ORF識(shí)別,用基于人工神經(jīng)網(wǎng)絡(luò)、隱馬爾科夫模型、及權(quán)重矩陣分析上述序列的信號(hào)肽及跨膜結(jié)構(gòu),從373條日本血吸蟲肝期童蟲體被蛋白序列中篩得14條具有ORF結(jié)構(gòu)及信號(hào)肽結(jié)構(gòu)的序列。 應(yīng)用免疫信息學(xué)技術(shù)進(jìn)行候選序列的Th細(xì)胞表位預(yù)測(cè)。應(yīng)用基于結(jié)合基序、矩陣法和人工神經(jīng)網(wǎng)絡(luò)等方法的多個(gè)服務(wù)器預(yù)測(cè)可與人源、鼠源MHCⅡ類分子結(jié)合的線性表位肽;應(yīng)用基于分子動(dòng)力學(xué)算法的受體配體嵌合空間結(jié)合模擬服務(wù)器,模擬肽與MHCⅡ分子的錨定情況,得到候選的Th細(xì)胞表位。結(jié)果預(yù)測(cè)得到了5個(gè)十五肽表位序列:NQFIRNLTLKIVLPE[源序列號(hào)AY813997;多萜基二磷酸寡聚糖蛋白葡萄糖基轉(zhuǎn)移酶類似物];SSTSQDFPGVCQLCV[AY815893;原鞘脂激活蛋白];SSLVISYSTVDRLVC[源序列號(hào)AY815893;原鞘脂激活蛋白];SRNLTVLRTNVALRL[源序列號(hào)AY811606;秀麗隱桿線蟲假象蛋白類似物]TVKFDKTVTCGGAYI[源序列號(hào)AAC00515;鈣網(wǎng)織蛋白家族]。 用人工合成多肽法(四分臂偶聯(lián)法合成到多聚賴氨酸骨架上)合成上述表位肽;MTT法改良技術(shù)CCK-8測(cè)定候選表位肽體外刺激免疫小鼠脾淋巴細(xì)胞增殖,結(jié)果初步鑒定獲得了1個(gè)童蟲期別蛋白候選Th細(xì)胞表位:SSLVISYSTVDRLVC[源序列號(hào)AY815893;原鞘脂激活蛋白]。 本研究采用免疫信息學(xué)理論預(yù)測(cè)技術(shù)結(jié)合體外試驗(yàn)檢測(cè)技術(shù),初步鑒定獲得了日本血吸蟲童蟲期別體被蛋白分子Th細(xì)胞表位,為進(jìn)一步篩選新的血吸蟲童蟲體被分子的保護(hù)性T細(xì)胞抗原奠定基礎(chǔ)。
[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，

本文編號(hào)：1851752