预防性人乳头瘤病毒病毒样颗粒疫苗的研究

发布时间：2018-04-18 03:37

本文选题：HPV + 病毒样颗粒　；参考：《吉林大学》2008年博士论文

【摘要】： 人乳头瘤病毒(Human Papillomavirus,HPV)有100多型别,根据与致癌性的关系分为高危型和低危型。高危型HPV与人类多种组织恶性肿瘤密切相关,尤其以HPV16、18、31、45、52和58型感染与宫颈癌的发生关系最为密切,其中HPV16占50%以上。本论文针对HPV开展了以下三方面的研究:(1)利用荧光素酶基因为报告基因与含有HPV6、16、18、31、45、52和58型L1、L2衣壳蛋白基因的真核表达质粒共转染293FT细胞,获得带有荧光素酶基因的七种假病毒,建立了针对七种亚型中和抗体的快速、灵敏、准确的检测方法。(2)利用HPV6、16、18、31、45、52、58等七种真核表达质粒免疫小鼠,考察七种亚型间中和抗体交叉保护情况,结果显示:HPV6、HPV16和HPV18免疫血清对HPV52与HPV58假病毒中和阻断率小于24.1%(p0.05);HPV58免疫血清对几种假病毒的中和阻断率分别为:HPV6(86.1%,p0.01)、HPV16(30.9%,p0.05)、HPV18(60.2%,p0.005)、HPV45(36.1%,p0.05)和HPV52(11.7%,p0.05)。(3)根据中国部分地区和世界范围内相关流行病学研究结果,同时结合中和抗体交叉保护的关系,确定HPV16和58型为中国地区疫苗设计主要型别。分别利用大肠杆菌表达系统和毕赤酵母(Pichia Pastoris)表达系统表达HPV16和58型的L1蛋白,其中原核表达体系表达的L1经过变性和复性过程,可形成L1五聚体;酵母表达体系分泌表达L1蛋白,可形成病毒样颗粒(Virus Like Particles,VLPs),直径约为50nm。免疫动物结果显示:HPV16和58型两种形式疫苗均可产生中和抗体;在免疫血清阻断假病毒感染的能力和疫苗免疫原性方面,病毒样颗粒疫苗优于五聚体疫苗。本论文研究结果为进一步研制适合中国和东亚地区的HPV预防性疫苗奠定基础。
[Abstract]:Human papillomavirus (HPV) has more than 100 types, which can be divided into high risk type and low risk type according to its relationship with carcinogenicity.High risk HPV is closely related to various malignant tumors of human tissues, especially HPV16 / 1831 / 455 / 52 and 58 / 58 infection, in which HPV16 accounts for more than 50% of the cases.In this paper, we carried out the following three studies on HPV: 1) by using luciferase gene and eukaryotic expression plasmids containing HPV6 / 16 / 1831H5 / 552 and 58 type L1 / L2 capsid protein gene, seven pseudoviruses with luciferase gene were obtained by co-transfection into 293FT cells.A rapid, sensitive and accurate detection method for seven subtypes of neutralizing antibodies was established.The results showed that the neutralization and blocking rates of HPV52 and HPV58 pseudoviruses in the two groups were lower than those in the control serum of 24. 1p0.05 and 24. 1p0. 05, respectively. The neutralization and blocking rates of several pseudoviruses were respectively:: HPV6 / 86.1and HPV161 / 30.9: p0.05HPV1860.2P0.005 / 36.01) and HPV5211.711.711.711.711.711.711.711.7/ p0.05.3.According to the results of epidemiological studies in some parts of China and the world, the neutralization and interdiction rates of HPV52 and HPV58 were found to be higher than that of HPV5 and HPV5, respectively, according to the results of epidemiological studies in some regions of China and in the world.At the same time, combined with the relationship of neutralizing antibody cross protection, HPV16 and 58 type were identified as the main types of vaccine design in China.HPV16 and 58 type L1 protein were expressed by E. coli expression system and Pichia pastoris expression system, respectively. The L1 expressed in prokaryotic expression system was denatured and renatured to form L1 pentamer.The yeast expression system secreted L1 protein and formed virus-like Like Particles-VLPsN with a diameter of about 50 nm.The results of immunized animals showed that both the two forms of vaccine: HPV16 and 58 could produce neutralizing antibodies, and that the virus-like granular vaccine was superior to the pentamer vaccine in the ability of blocking the infection of pseudovirus and the immunogenicity of the vaccine.The results of this paper lay a foundation for the further development of HPV prophylactic vaccine suitable for China and East Asia.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2008
【分类号】：R392.11

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