缺失突變HCV核蛋白DNA免疫效果的初步評(píng)價(jià)
發(fā)布時(shí)間:2018-06-19 03:57
本文選題:丙型肝炎病毒 + 核蛋白; 參考:《第四軍醫(yī)大學(xué)》2005年博士論文
【摘要】:丙型肝炎病毒(hepatitis C virus,HCV)感染已成為困擾全球人口健康的一大問題,超過1.7億人有HCV感染。HCV感染后85%將發(fā)展為慢性,約20%最終發(fā)展為終末期肝病。到目前為止尚無切實(shí)有效的預(yù)防和治療方法。因此,迫切需要開發(fā)切實(shí)有效的疫苗來預(yù)防感染和尋求新的治療途徑。 長期以來,HCV核蛋白作為抗HCV疫苗的備選抗原一直受到關(guān)注。DNA免疫研究揭示編碼HCV核蛋白的DNA疫苗能夠誘導(dǎo)特異性抗體產(chǎn)生和Th1類細(xì)胞免疫反應(yīng),然而由于完整的核蛋白基因疫苗免疫原性十分有限,促使研究者尋求多種策略進(jìn)行優(yōu)化,但仍然未達(dá)到理想的效果。核蛋白DNA疫苗在細(xì)胞內(nèi)表達(dá)量不足和核蛋白本身對(duì)免疫功能的抑制作用可能是影響提高核蛋白特異性免疫的兩個(gè)重要因素。本研究著眼于對(duì)核蛋白自身結(jié)構(gòu)的改造,試圖通過提高核蛋白在真核細(xì)胞內(nèi)的表達(dá)水平、去除影響免疫細(xì)胞對(duì)核蛋白有效識(shí)別的肽段,達(dá)到增強(qiáng)核蛋白特異性免疫反應(yīng)的目的。 實(shí)驗(yàn)方法和結(jié)果: 1.通過RT-PCR方法從西安地區(qū)慢性丙型肝炎患者血清中擴(kuò)增了HCV C區(qū)及部分E1區(qū)cDNA,將其插入pGEM-Teasy載體中,采用酶切
[Abstract]:Hepatitis C virus (HCV) infection has become a major health problem in the world. More than 170 million people have HCV infection. 85% of them will become chronic after infection, and about 20% will eventually develop into end-stage liver disease. So far, there are no effective prevention and treatment methods. Therefore, there is an urgent need to develop effective vaccines to prevent infection and seek new ways of treatment. For a long time, HCV nucleoprotein as an alternative antigen of anti-HCV vaccine has been concerned. DNA immunological studies have revealed that the DNA vaccine encoding HCV nucleoprotein can induce specific antibody production and Th1 cell immune response. However, due to the limited immunogenicity of the complete nucleoprotein gene vaccine, many strategies have been sought for optimization, but the desired results have not been achieved. The insufficient expression of nucleoprotein DNA vaccine in cells and the inhibition of nuclear protein itself on immune function may be two important factors affecting the enhancement of specific immunity of nucleoprotein. The aim of this study is to improve the expression level of nucleoprotein in eukaryotic cells and to remove the peptides that affect the effective recognition of nucleoprotein in order to enhance the specific immune response of nucleoprotein. Experimental methods and results: 1. The HCV C region and some E1 region cDNAs were amplified by RT-PCR from the serum of patients with chronic hepatitis C in Xi'an area. They were inserted into pGEM-T easy vector and digested by enzyme.
【學(xué)位授予單位】:第四軍醫(yī)大學(xué)
【學(xué)位級(jí)別】:博士
【學(xué)位授予年份】:2005
【分類號(hào)】:R392
【引證文獻(xiàn)】
相關(guān)碩士學(xué)位論文 前1條
1 沈才飛;不同缺失突變的丙型肝炎病毒核蛋白在大腸桿菌中的表達(dá)[D];第四軍醫(yī)大學(xué);2006年
,本文編號(hào):2038319
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