白色念珠菌热休克蛋白90核酸疫苗的构建及其免疫学活性研究

发布时间：2018-05-31 22:22

本文选题：核酸疫苗 + Hsp90　；参考：《东北师范大学》2011年硕士论文

【摘要】：核酸疫苗(DNA vaccine)是指将含有编码某种抗原蛋白的外源基因序列插入到质粒载体中,该重组质粒就称为核酸疫苗。核酸疫苗又称为基因疫苗。核酸疫苗是继病原体疫苗、亚单位疫苗之后出现的第三代疫苗,与传统的疫苗相比具有多价、高效、价廉、安全等许多优势。白色念珠菌(Candida albicans)是一种寄生在动物和人的口腔和胃肠道的条件致病菌,它可以在机体免疫力下降和正常菌群失调时作为机会病原体,引起机体的浅层感染(也称粘膜性感染)和系统性感染(也称深层感染或全身性感染)。系统性白色念珠菌感染危害巨大,据科学家统计,在使用了抗真菌药物后,该病的死亡率仍高达70%以上。随着发病率和致死率的不断增加,越来越多的科学工作者投入到白色念珠菌免疫学机制的研究中。英国曼彻斯特医学院的Matthews等人长期从事白色念珠菌免疫学方面的研究,他们发现在系统性白色念珠菌感染中热休克蛋白90(Hsp90)起着主要的病理学作用。并确定了Hsp90的抗原表位图谱,发现了2个能够与病人血清起反应的常见表位,即表位C和H,表位C是一种保守表位,能够与所有病人的血清起反应;表位H是白色念珠菌所特有的一种表位,它能够被大约50%的病人血清识别。本实验选择了Hsp90全基因序列中含有编码C、H表位的一段DNA序列,构建了核酸疫苗pcDNA3.1/Hsp90。通过肌肉注射后电转(electroporation)的方式免疫小鼠, Western blot结果表明:经热休克蛋白90重组质粒(pcDNA3.1/Hsp90)免疫小鼠抗血清中产生了抗热休克蛋白90重组蛋白(R-Hsp90)特异性抗体; ELISA结果表明:经pcDNA3.1/Hsp90组免疫后小鼠抗血清中抗R-Hsp90特异性抗体IgG的抗体含量,与pcDNA3.1组和PBS组比较得到了显著提高(P0.01)。本实验为进一步研究白色念珠菌核酸疫苗奠定了坚实基础。
[Abstract]:Nucleic acid vaccine (DNA accine) is a recombinant plasmid called nucleic acid vaccine by inserting the foreign gene sequence which encodes a certain antigen protein into the plasmid vector. Nucleic acid vaccines are also called genetic vaccines. Nucleic acid vaccine is the third generation vaccine after pathogen vaccine and subunit vaccine. Compared with traditional vaccine, nucleic acid vaccine has many advantages, such as multivalent, high efficiency, low price, safety and so on. Candida albicansis, a opportunistic pathogen that parasitizes the oral and gastrointestinal tract of animals and humans, can act as an opportunistic pathogen when the body's immunity declines and its normal flora is dysfunctional. Causes superficial infection (also known as mucosal infection) and systemic infection (also known as deep infection or systemic infection). Systemic Candida albicans infection is harmful, according to scientists statistics, after the use of antifungal drugs, the death rate is still as high as 70%. With the increasing incidence and fatality rate, more and more scientists are involved in the study of immunological mechanism of Candida albicans. Matthews et al., from Manchester Medical College in England, have long been involved in the immunological study of Candida albicans. They have found that heat shock protein 90 (Hsp90) plays a major role in the pathogenesis of systemic Candida albicans infection. The antigenic epitope map of Hsp90 was determined. Two common epitopes, epitopes C and H, were found to react with the sera of patients. Epitope C was a conserved epitope and could react with the sera of all patients. Epitope H, a specific epitope of Candida albicans, can be recognized by about 50% of patient serum. In this study, we selected a sequence of DNA encoding CnH epitope in the whole Hsp90 gene sequence, and constructed pcDNA3.1 / Hsp90 nucleic acid vaccine. , Western blot was immunized by electroporation of mice after intramuscular injection. The results showed that the anti-heat shock protein 90 recombinant protein R-Hsp90 antibody was produced in the antiserum of mice immunized with recombinant plasmid pcDNA3.1 / Hsp90. Ming: the antibody content of anti R-Hsp90 specific antibody IgG in the antiserum of mice immunized with pcDNA3.1/Hsp90 group, Compared with pcDNA3.1 group and PBS group, there was a significant increase in P0.01. This study laid a solid foundation for further study of Candida albicans nucleic acid vaccine.
【学位授予单位】：东北师范大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392

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