發(fā)布時(shí)間：2018-04-18 03:37

  本文選題：HPV + 病毒樣顆粒��； 參考：《吉林大學(xué)》2008年博士論文

【摘要】： 人乳頭瘤病毒(Human Papillomavirus,HPV)有100多型別,根據(jù)與致癌性的關(guān)系分為高危型和低危型。高危型HPV與人類(lèi)多種組織惡性腫瘤密切相關(guān),尤其以HPV16、18、31、45、52和58型感染與宮頸癌的發(fā)生關(guān)系最為密切,其中HPV16占50%以上。 本論文針對(duì)HPV開(kāi)展了以下三方面的研究:(1)利用熒光素酶基因?yàn)閳?bào)告基因與含有HPV6、16、18、31、45、52和58型L1、L2衣殼蛋白基因的真核表達(dá)質(zhì)粒共轉(zhuǎn)染293FT細(xì)胞,獲得帶有熒光素酶基因的七種假病毒,建立了針對(duì)七種亞型中和抗體的快速、靈敏、準(zhǔn)確的檢測(cè)方法。(2)利用HPV6、16、18、31、45、52、58等七種真核表達(dá)質(zhì)粒免疫小鼠,考察七種亞型間中和抗體交叉保護(hù)情況,結(jié)果顯示:HPV6、HPV16和HPV18免疫血清對(duì)HPV52與HPV58假病毒中和阻斷率小于24.1%(p0.05);HPV58免疫血清對(duì)幾種假病毒的中和阻斷率分別為: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)根據(jù)中國(guó)部分地區(qū)和世界范圍內(nèi)相關(guān)流行病學(xué)研究結(jié)果,同時(shí)結(jié)合中和抗體交叉保護(hù)的關(guān)系,確定HPV16和58型為中國(guó)地區(qū)疫苗設(shè)計(jì)主要型別。分別利用大腸桿菌表達(dá)系統(tǒng)和畢赤酵母(Pichia Pastoris)表達(dá)系統(tǒng)表達(dá)HPV16和58型的L1蛋白,其中原核表達(dá)體系表達(dá)的L1經(jīng)過(guò)變性和復(fù)性過(guò)程,可形成L1五聚體;酵母表達(dá)體系分泌表達(dá)L1蛋白,可形成病毒樣顆粒(Virus Like Particles,VLPs),直徑約為50nm。免疫動(dòng)物結(jié)果顯示:HPV16和58型兩種形式疫苗均可產(chǎn)生中和抗體;在免疫血清阻斷假病毒感染的能力和疫苗免疫原性方面,病毒樣顆粒疫苗優(yōu)于五聚體疫苗。 本論文研究結(jié)果為進(jìn)一步研制適合中國(guó)和東亞地區(qū)的HPV預(yù)防性疫苗奠定基礎(chǔ)。
[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.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2008
【分類(lèi)號(hào)】：R392.11

【共引文獻(xiàn)】

相關(guān)期刊論文 前1條

1 田厚文;任皎;黃薇;范江濤;趙莉;阮力;;表達(dá)人乳頭瘤病毒16型E6/E7融合蛋白的非復(fù)制型重組痘苗病毒的構(gòu)建及免疫效果觀察[J];病毒學(xué)報(bào);2006年05期

相關(guān)博士學(xué)位論文 前6條

1 許雪梅;人乳頭瘤病毒16型E6E7基因的克隆、修飾及與共激活分子B7-1協(xié)同誘導(dǎo)特異性免疫反應(yīng)的研究[D];中國(guó)協(xié)和醫(yī)科大學(xué);2002年

2 尹銳;人乳頭瘤病毒16E7 HLA-A_2限制性CTL表位的穿膜肽疫苗設(shè)計(jì)及體外免疫學(xué)研究[D];第三軍醫(yī)大學(xué);2006年

3 劉方;一、間接免疫熒光技術(shù)檢測(cè)尖銳濕疣中的人乳頭瘤病毒 二、HPV6b L1 DNA疫苗的構(gòu)建及免疫活性檢測(cè)[D];中國(guó)協(xié)和醫(yī)科大學(xué);2004年

4 周曉波;人乳頭瘤病毒（HPV）在中國(guó)食管癌中的檢測(cè)以及HPV16 E7作用機(jī)制的研究[D];中國(guó)協(xié)和醫(yī)科大學(xué);2003年

5 左亞剛;安全高效HPV 16E7 DNA疫苗的構(gòu)建及免疫原性分析[D];中國(guó)協(xié)和醫(yī)科大學(xué);2003年

6 相文忠;HPV11-E6/E7與人IFNα-2b融合基因表達(dá)質(zhì)粒的構(gòu)建、表達(dá)與實(shí)驗(yàn)免疫研究[D];南京醫(yī)科大學(xué);2006年

相關(guān)碩士學(xué)位論文 前3條

1 胡云峰;復(fù)發(fā)性尖銳濕疣HPV6 L1基因原核表達(dá)質(zhì)粒構(gòu)建及序列分析[D];暨南大學(xué);2005年

2 李義飛;MCM5、MCM7在宮頸癌及癌前病變中的表達(dá)及與HPV16感染的關(guān)系[D];河北醫(yī)科大學(xué);2006年

3 殷董;人乳頭瘤病毒6bL1雙順?lè)醋虞d體的構(gòu)建及其在哺乳動(dòng)物細(xì)胞內(nèi)的表達(dá)[D];暨南大學(xué);2007年

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本文編號(hào)：1766659