周期型马来丝虫Bm-GAPDH核酸疫苗在小鼠体内的免疫应答及Bm-CPI基因克隆的研究
发布时间：2018-10-15 19:42
【摘要】：丝虫病是一种严重危害人类健康的传染病,长期使用化学药物,使丝虫和蚊媒抗药性增强。研究和发展价格低廉、安全高效的丝虫疫苗是未来丝虫病防治工作中极有潜力的重要课题。近年来发展起来的核酸疫苗以其制备简便、实用高效等优点而备受重视,具有广阔的应用前景。现有的资料表明3-磷酸甘油醛脱氢酶(glyceraldehyde-3-phosphate dehydrogenase, GAPDH)在寄生蠕虫和原虫中,可诱导实验动物产生保护性免疫应答,是一种重要的疫苗候选抗原。本研究成功构建了周期型马来丝虫核酸疫苗pcDNA3.1/Bm-gapdh。为了提高免疫效应,用免疫刺激序列CpG作为佐剂共同免疫BALB/c小鼠,用RT-PCR、MTT、ELISA检测GAPDH的免疫效应。结果表明,核酸疫苗pcDNA3.1/Bm-gapdh具有抗丝虫病候选DNA疫苗分子的潜力,增加佐剂后免疫效应有所增强。为了下一步的疫苗设计作前期准备,本实验还构建了周期型马来丝虫半胱氨酸蛋白酶抑制剂(cysteine protease inhibitor,CPI)的重组质粒pGEM-T/Bm-cpi,并送基因公司测序。再根据测序结果,推测其氨基酸序列,然后利用相关的软件预测它的B细胞表位。
[Abstract]:Filariasis is an infectious disease that seriously endangers human health. Long-term use of chemical drugs enhances resistance to filariasis and mosquito vectors. The research and development of low cost, safe and efficient filariasis vaccine is an important topic with great potential in filariasis control in the future. The nucleic acid vaccine which has been developed in recent years has attracted much attention because of its advantages of simple preparation, practical efficiency and so on, and has a broad application prospect. The available data show that glyceraldehyde-3-phosphate dehydrogenase, GAPDH) can induce protective immune response of experimental animals in parasitic worms and protozoa and is an important vaccine candidate antigen. In this study, we successfully constructed the pcDNA3.1/Bm-gapdh. of periodic filariasis nucleic acid vaccine. In order to improve the immune effect, BALB/c mice were immunized with CpG as adjuvant, and the immune effect of GAPDH was detected by RT-PCR,MTT,ELISA. The results showed that the nucleic acid vaccine pcDNA3.1/Bm-gapdh had the potential to be a candidate DNA vaccine against filariasis, and the immune effect was enhanced after increasing the adjuvant. In order to prepare the vaccine for the next step, the recombinant plasmid pGEM-T/Bm-cpi, of cysteine protease inhibitor (cysteine protease inhibitor,CPI (Cysteine protease inhibitor) of filariasis malayi was constructed and sequenced. The amino acid sequence was deduced according to the results of sequencing, and then the B cell epitopes were predicted by the relevant software.
【学位授予单位】：南通大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392
[Abstract]:Filariasis is an infectious disease that seriously endangers human health. Long-term use of chemical drugs enhances resistance to filariasis and mosquito vectors. The research and development of low cost, safe and efficient filariasis vaccine is an important topic with great potential in filariasis control in the future. The nucleic acid vaccine which has been developed in recent years has attracted much attention because of its advantages of simple preparation, practical efficiency and so on, and has a broad application prospect. The available data show that glyceraldehyde-3-phosphate dehydrogenase, GAPDH) can induce protective immune response of experimental animals in parasitic worms and protozoa and is an important vaccine candidate antigen. In this study, we successfully constructed the pcDNA3.1/Bm-gapdh. of periodic filariasis nucleic acid vaccine. In order to improve the immune effect, BALB/c mice were immunized with CpG as adjuvant, and the immune effect of GAPDH was detected by RT-PCR,MTT,ELISA. The results showed that the nucleic acid vaccine pcDNA3.1/Bm-gapdh had the potential to be a candidate DNA vaccine against filariasis, and the immune effect was enhanced after increasing the adjuvant. In order to prepare the vaccine for the next step, the recombinant plasmid pGEM-T/Bm-cpi, of cysteine protease inhibitor (cysteine protease inhibitor,CPI (Cysteine protease inhibitor) of filariasis malayi was constructed and sequenced. The amino acid sequence was deduced according to the results of sequencing, and then the B cell epitopes were predicted by the relevant software.
【学位授予单位】：南通大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392
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