本文選題：單純皰疹病毒Ⅰ型 + ul7��； 參考：《北京協(xié)和醫(yī)學(xué)院》2017年博士論文

【摘要】：人類皰疹病毒作為一個(gè)由8個(gè)成員組成的DNA病毒家族,在全球不同人群中均具有較高的感染率,尤其是其中的HSV-1和HSV-2等病原所引起的包括口唇皰疹、生殖器官皰疹以及皰疹性腦炎等常見(jiàn)感染,已成為青少年人群中的重要傳染性疾病,并受到公共衛(wèi)生領(lǐng)域的廣泛關(guān)注。因此,針對(duì)這一病原的相關(guān)臨床治療和預(yù)防性制品的研究已經(jīng)得到了多方面的推進(jìn)。迄今為止,盡管阿昔洛韋類藥物已經(jīng)在臨床上得到了廣泛的應(yīng)用,但有關(guān)HSV-1和HSV-2的多個(gè)疫苗臨床研究均未提供具有應(yīng)用意義的結(jié)果�；贖SV-1和HSV-2感染病理的特征探索合適的疫苗形式以誘導(dǎo)有效的臨床保護(hù)效率顯然是目前一個(gè)重要的研究方向。我們前期關(guān)于HSV-1與宿主相互作用的分子相關(guān)研究以及HSV-1相關(guān)突變株的構(gòu)建技術(shù)的研究,為進(jìn)一步針對(duì)HSV-1衣被蛋白在病毒生物學(xué)特性及其毒力表型變化中的作用分析提供了初步的資料。在此基礎(chǔ)上,本文的主要工作描述了一個(gè)具有轉(zhuǎn)錄調(diào)控作用的HSV-1衣被蛋白UL7突變后對(duì)HSV-1毒力表型的影響,及其可能機(jī)理的研究過(guò)程�；谶@一突變毒株的生物學(xué)特性,我們?cè)谄浠A(chǔ)上完成了 HSV-1復(fù)制過(guò)程中的重要調(diào)控分子Vhs編碼基因u141和與潛伏感染的建立和維持密切相關(guān)的LAT基因的突變,構(gòu)建了突變毒株M2和M3,并對(duì)三株毒株的生物學(xué)特性及致病性進(jìn)行了分析。最后,我們以突變毒株M3為研究重點(diǎn),評(píng)價(jià)了其在小鼠模型上的免疫原性,為HSV-1減毒病毒的研究提供了初步的基礎(chǔ)。在第一部分工作中,針對(duì)UL7突變的毒株M1的分析表明,突變毒株M1表現(xiàn)出細(xì)胞水平的低增殖能力以及動(dòng)物水平的急性期感染能力的下降,進(jìn)一步的體外實(shí)驗(yàn)還表明UL7蛋白可能通過(guò)參與染色質(zhì)化α-4基因而起到調(diào)控其轉(zhuǎn)錄激活的作用。在第二部分工作中,突變毒株M1、M2和M3的生物學(xué)特性分析結(jié)果表明,與野生型毒株相比,突變毒株M3在細(xì)胞中的增殖能力顯著降低,在小鼠體內(nèi)的致病能力和潛伏能力均顯著降低,提示了 M3毒株的減毒特性�；谇皟刹糠值难芯抠Y料,我們將突變毒株M3作為減毒活疫苗候選株免疫小鼠,并在免疫后1個(gè)月和2個(gè)月時(shí)分別予以野生型毒株滴鼻感染。隨后的臨床癥狀觀察、免疫學(xué)、組織病理學(xué)和病毒載量測(cè)定結(jié)果顯示,突變毒株M3免疫后誘導(dǎo)的以中和抗體和CD8+T細(xì)胞免疫反應(yīng)為特征的免疫反應(yīng)在針對(duì)野生型毒株攻擊情況下,能夠有效控制野生型毒株感染引起的多種臨床癥狀,抑制野生型毒株在小鼠體內(nèi)的增殖,顯著減輕野生型毒株感染引起的病理?yè)p傷,限制野生型毒株在神經(jīng)系統(tǒng)的潛伏感染過(guò)程。這些結(jié)果均表明了突變毒株M3可作為減毒活疫苗候選株的潛在可能。
[Abstract]:As a family of eight members of DNA virus, human herpesvirus has a high infection rate in different populations around the world, especially HSV-1, HSV-2 and other pathogens, including oral herpes.Herpes genitalis and herpes encephalitis and other common infections have become an important infectious disease in the adolescent population and have received extensive attention in the field of public health.Therefore, the research on clinical treatment and prophylaxis of this pathogen has been advanced in many aspects.So far, although acyclovir drugs have been widely used in clinical practice, many clinical studies on HSV-1 and HSV-2 have not provided significant results.Based on the pathological features of HSV-1 and HSV-2 infection, it is an important research direction to explore the appropriate vaccine form to induce effective clinical protection efficiency.We have studied the molecular correlation of HSV-1 interaction with host and the construction of HSV-1 related mutants.It provides preliminary data for the analysis of the role of HSV-1 coat protein in the changes of virus biological characteristics and virulence phenotypes.On this basis, the main work described a transcriptional regulation of HSV-1 coat protein UL7 mutation after the impact of HSV-1 virulence phenotype, and the possible mechanism of the study process.Based on the biological characteristics of the mutant strain, we have completed the mutation of the Vhs encoding gene U141, an important regulatory molecule in the process of HSV-1 replication, and the LAT gene closely related to the establishment and maintenance of latent infection.Mutants M2 and M3 were constructed and their biological characteristics and pathogenicity were analyzed.Finally, we evaluated the immunogenicity of mutant strain M3 in mouse model and provided a preliminary basis for the study of HSV-1 attenuated virus.In the first part of the work, the analysis of M1 strain with UL7 mutation showed that mutant M1 showed low proliferation ability at cell level and decreased infection ability at animal level in acute stage.Further in vitro experiments indicated that UL7 protein may regulate the transcriptional activation by participating in chromatin 偽 -4 gene.In the second part of the work, the biological characteristics of mutant strain M1M2 and M3 were analyzed. The results showed that the proliferation of mutant strain M3 was significantly lower than that of wild-type strain.The pathogenicity and latent ability in mice were significantly decreased, indicating the detoxification characteristics of M _ 3 strain.Based on the first two parts of the study, we immunized mice with mutant strain M3 as a candidate strain of attenuated live vaccine, and were inoculated with wild-type virus strain drip nasal infection at 1 month and 2 months after immunization, respectively.Subsequent clinical observation, immunology, histopathology, and viral load measurements showed that,The immune response induced by mutant strain M3, characterized by neutralizing antibody and CD8 T cell immune response, can effectively control the clinical symptoms caused by wild-type virus infection under the attack of wild-type strain.Inhibiting the proliferation of wild type virus strain in mice, reducing the pathological damage caused by wild type virus strain infection, limiting the latent infection process of wild type virus strain in nervous system.These results suggest that mutant strain M _ 3 is a potential candidate for attenuated live vaccine.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：R392

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