發(fā)布時間：2019-07-04 13:46

【摘要】： 戊型肝炎病毒(HEV)是戊型肝炎的病原體,隨著HEV流行病學研究的深入,發(fā)現(xiàn)HEV的4個主要基因型存在著明顯的易感宿主差異:基因1型(HEV-1)和基因2型(HEV-2)僅分離于人類,能導致大規(guī)模的戊肝爆發(fā)流行,實驗動物目前只成功感染非人靈長類,而基因3型(HEV-3)和基因4型(HEV-4)人畜共患,僅見于小規(guī)模流行和臨床散發(fā)。我們將HEV分為兩類:包括HEV-1/2的H(Human)類和包括HEV-3/4的Z(Zoonosis)類。 首先我們通過Western blot、體外捕獲PCR、ELISA阻斷實驗及合成的多肽庫對實驗室制備的多株抗HEV單抗的識別表位進行系統(tǒng)的研究,結果發(fā)現(xiàn)12株線性單抗識別的位都位于ORF2 aa408-458之間,17株構象型單抗識別表位都定位于ORF2 aa459-606之間,其中15株的識別表位位于天然病毒表面。 現(xiàn)有研究結果表明HEV的主要中和表位區(qū)域集中于ORF2的aa459-606之間,并且也是主要介導HEV與嗜性細胞吸附的區(qū)域。本研究通過比較這兩類HEV ORF2 aa368-606區(qū)段,發(fā)現(xiàn)存在4個類保守的差異位點,均位于HEV的主要中和表位區(qū)域,分別是Ser483Thr、Val492Met、Ser497Thr和Ala599Gly。以能形成類病毒顆粒的HEV 239(ORF2 aa368-606)為基礎,對這四個位點進行定點替換突變,并以這15株能夠捕獲HEV-1和/或HEV-4的單克隆抗體比較各種突變體的免疫反應性,結果表明僅aa497的差異造成了這兩類HEV中和表位構象的部分差異,提示aa497及其相關的病毒表面結構差異可能在H類和Z類HEV宿主選擇中扮演重要角色。
文內圖片：
圖片說明：HEV基因組Fig1GenomicorganizationofHEV(A)ingenotype1-3and(B)ingenotype4
[Abstract]:Hepatitis E virus (HEV) is the pathogen of hepatitis E. with the deepening of HEV epidemiological research, it is found that there are obvious susceptible host differences in the four main genotypes of HEV: gene type 1 (HEV-1) and gene type 2 (HEV-2) are only isolated from human beings, which can lead to large-scale outbreak of hepatitis E. At present, experimental animals are only successfully infected with non-human primates. Gene 3 (HEV-3) and gene 4 (HEV-4) zoonosis are only found in small-scale epidemic and clinical dissemination. We divide HEV into two categories: the H (Human) class of HEV-1/2 and the Z (Zoonosis) class of HEV-3/4. Firstly, we systematically studied the recognition epitopes of several strains of anti-HEV monoclonal antibodies prepared in laboratory by Western blot, capture PCR,ELISA blocking experiment in vitro and the synthetic polypeptide library. The results showed that the recognition epitopes of 12 strains of linear monoclonal antibodies were located between ORF2 aa408-458 and 17 strains of constructed monoclonal antibodies were located between ORF2 aa459-606, and 15 of them were located on the surface of natural viruses. The existing results show that the main neutralizing epitope region of HEV is concentrated between the aa459-606 of ORF2, and it is also the region that mediates the adsorption of HEV with sex-like cells. In this study, by comparing the two types of HEV ORF2 aa368-606 regions, it was found that there were four conserved difference sites, all of which were located in the main neutralizing epitope regions of HEV, Ser483Thr,Val492Met,Ser497Thr and Ala599Gly., respectively. The site-directed substitution mutations of these four loci were carried out on the basis of HEV-239( ORF2 aa368-606), which can form viral particles, and the immunoreactivity of various mutants was compared with the 15 monoclonal antibodies that could capture HEV-1 and / or HEV-4. The results showed that only the difference of aa497 resulted in some differences in the conformational patterns of neutralizing epitopes between the two types of HEV. It is suggested that the surface structure difference of aa497 and its related viruses may play an important role in the host selection of class H and Z HEV.
【學位授予單位】：廈門大學
【學位級別】：碩士
【學位授予年份】：2007
【分類號】：R392

【參考文獻】

相關期刊論文 前10條

1 顧穎,葛勝祥,黃果勇,李少偉,朱子恒,何志強,陳毅歆,王穎彬,張軍,夏寧邵;戊型肝炎病毒中和性單克隆抗體的鑒定[J];病毒學報;2003年03期

2 張軍,顧穎,歐山海,王穎彬,葉祥忠,林鑒,葛勝祥,夏寧邵;戊型肝炎病毒衣殼蛋白中和表位間的構象誘導[J];病毒學報;2004年02期

3 葛勝祥,郭清順,李少偉,張軍,夏寧邵;基因Ⅰ、Ⅳ型戊型肝炎病毒高靈敏度通用引物的設計和初步應用[J];病毒學報;2005年03期

4 何水珍;鄭子崢;吳婷;謝明輝;苗季;張軍;夏寧邵;;戊型肝炎病毒細胞吸附模型的建立及病毒吸附區(qū)域初步研究[J];病毒學報;2006年06期

5 莊輝,劉崇柏,曹學義,王光明;戊型肝炎的病原學及流行病學研究[J];北京醫(yī)科大學學報;1992年04期

6 李少偉,張軍,何志強,葛勝祥,顧穎,林鑒,劉如石,夏寧邵;大腸桿菌表達的戊型肝炎病毒ORF2片段的聚合現(xiàn)象研究[J];生物工程學報;2002年04期

7 李少偉,何志強,王穎彬,陳毅歆,劉如石,林鑒,顧穎,張軍,夏寧邵;戊型肝炎病毒衣殼蛋白同源二聚體的相互作用結構域[J];生物工程學報;2004年01期

8 夏小兵,黃如統(tǒng),李德榮;戊型肝炎病毒亞基因組RNA的研究[J];微生物學報;2000年06期

9 葛勝祥,張軍,黃果勇,逄淑強,周開姣,夏寧邵;大腸桿菌表達的戊型肝炎病毒ORF2多肽對恒河猴的免疫保護研究[J];微生物學報;2003年01期

10 李奎,莊輝,朱萬孚;從中國 14個城市戊型肝炎病人血清中檢測到的戊型肝炎病毒部分核苷酸序列分析(英文)[J];Chinese Medical Journal;2002年07期

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