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  本文關(guān)鍵詞： 乙型 丙型 肝炎 慢性 IP-10 NK細胞　出處：《吉林大學(xué)》2013年博士論文　論文類型：學(xué)位論文

【摘要】：乙型肝炎病毒(Hepatitis B virus，HBV)和丙型肝炎病毒(Hepatitis C virus，HCV)感染是導(dǎo)致肝炎、肝硬化、肝癌非常重要的危險因素。目前我國大概有9300萬的乙肝感染者和600~1000萬丙肝感染者。宿主的免疫因素在乙型肝炎及丙型肝炎的易感性、疾病的發(fā)生發(fā)展、對治療的反應(yīng)等方面發(fā)揮著重要作用。宿主固有免疫中的NK細胞在慢性乙型肝炎的發(fā)生發(fā)展及趨化因子IP-10的血漿學(xué)表達及其單核苷酸多態(tài)性在慢性丙型肝炎的易感性及對抗病毒治療應(yīng)答均發(fā)揮著重要的作用。本實驗將闡述宿主免疫因素對慢性乙型肝炎及慢性丙型肝炎的發(fā)病機制及對抗病毒治療療效的影響。 趨化因子是一類具有化學(xué)趨化活性的細胞因子，在炎癥反應(yīng)中起重要作用。大量研究證實：趨化因子的異常表達與病毒性肝炎疾病的進程密切相關(guān)。高表達的趨化因子可招募大量效應(yīng)淋巴細胞如細胞毒性T淋巴細胞至肝內(nèi)以清除病毒感染的肝細胞，但同時由于趨化因子的過度釋放，，導(dǎo)致炎癥反應(yīng)擴大，加劇了肝細胞損傷。因此明確相關(guān)趨化因子在病毒性肝炎中的作用機制尤為重要。 IP-10是1985年發(fā)現(xiàn)的趨化因子CXC亞族成員，在病毒性肝炎的發(fā)生發(fā)展及肝臟的免疫損傷中扮演著重要角色。多項研究顯示，在慢性丙型肝炎患者肝臟及血漿中檢測到高水平IP-10的表達。其高表達與肝臟炎癥活動程度和纖維化密切相關(guān)。目前，IP-10的單核苷酸多態(tài)性與血漿學(xué)表達與HCV感染后的結(jié)局及干擾素治療后應(yīng)答的關(guān)系還不清楚。 NK細胞是天然免疫細胞，在抗病毒免疫中發(fā)揮重要作用。NK細胞可以直接殺傷病毒感染的細胞并能間接通過抗體介導(dǎo)發(fā)揮細胞介導(dǎo)的細胞毒作用。NK細胞的功能取決于其活化性受體和抑制性受體與不同種類細胞表面的配體相結(jié)合。這些配體包括典型的和非典型的MHC I類抗原，MHC樣蛋白和其他各種自身和病毒衍生的分子。它們可能組成形表達或表達于病毒感染的細胞表面。 在HBV感染的過程及應(yīng)用核苷酸類似物抗病毒治療過程中NK細胞所起到的作用及變化鮮有報道。 因此本研究第一部分采用質(zhì)譜技術(shù)和酶聯(lián)免疫吸附測定法等技術(shù)：檢測慢性丙型肝炎患者、感染慢性丙型肝炎病毒后自發(fā)清除者及健康對照者中血漿IP-10表達水平的差異；觀察慢性丙型肝炎患者在干擾素聯(lián)合利巴韋林抗病毒治療前、抗病毒治療中及抗病毒治療后患者血漿IP-10表達的變化；結(jié)合慢性丙型肝炎患者抗病毒治療的療效，找到能預(yù)測療效的最佳的IP-10表達閾值；在慢性丙型肝炎患者、自發(fā)清除者及健康對照者中對IP-10的SNP位點rs3921；rs8878；rs4859584；rs4241578；rs4859588；rs56061981；rs74810361；rs4256246進行測序分析，分析IP-10不同的SNP位點的不同表型與慢性丙型肝炎感染及抗病毒治療后療效的關(guān)系。 第一部分實驗結(jié)果如下：HCV自發(fā)清除者血漿IP-10表達比慢性丙型肝炎患者及健康對照者低，同時健康對照者比慢性丙型肝炎患者血漿IP-10表達低；用于治療慢性丙型肝炎患者的干擾素和利巴韋林會誘使血漿IP-10表達增高，但治療結(jié)束后24周時血漿IP-10表達比未治療前明顯降低；在干擾素聯(lián)合利巴韋林抗病毒治療前基線IP-10表達小于600pg/ml是患者容易獲得完全早期病毒學(xué)應(yīng)答的獨立預(yù)測因素；在干擾素聯(lián)合利巴韋林抗病毒治療后2周時IP-10表達小于540pg/ml是患者容易獲得早期病毒學(xué)應(yīng)答的獨立預(yù)測因素；IP-10基因中所檢測的8個SNP位點與慢性丙型肝炎病毒易感性無關(guān)，與抗病毒療效無關(guān)。 第二部分采用流式細胞技術(shù)觀察替諾福韋組及阿德福韋酯組患者的抗病毒療效；檢測慢性乙型肝炎患者外周血中NK細胞數(shù)量及受體表達與正常對照者之間的差異；觀察替諾福韋組及阿德福韋酯組在抗病毒治療過程中NK細胞受體表達的變化情況，并比較兩者間的差異；找到可能預(yù)測慢性乙型肝炎患者抗病毒療效的NK細胞受體。 第二部分實驗結(jié)果如下：替諾福韋和阿德福韋酯治療的乙肝患者具有不同的抗病毒療效，替諾福韋抗病毒療效優(yōu)于阿德福韋酯；慢性乙型肝炎患者肝臟損傷與NK細胞受體表達的改變有關(guān)；應(yīng)用替諾福韋或阿德福韋酯抗病毒治療可以降低慢性乙型肝炎患者NK細胞的抑制性受體NKG2A~+和KIR2DL3~+的表達；NKG2A~+和KIR2DL3~+受體的表達可能是臨床上評價慢性乙型肝炎抗病毒療效的預(yù)測因子。 綜上所述，IP-10在慢性丙型肝炎感染及抗病毒治療過程中發(fā)揮著重要的作用，并且IP-10表達的高低可直接預(yù)測患者抗病毒的早期療效；NK細胞在慢性乙型肝炎感染過程中發(fā)揮著重要作用，并且NKG2A~+和KIR2DL3~+受體的表達可能是臨床上評價慢性乙型肝炎抗病毒療效的預(yù)測因子。
[Abstract]:Hepatitis B virus (Hepatitis B, virus, HBV) and hepatitis C virus (Hepatitis C, virus, HCV) infection is the leading cause of hepatitis, liver cirrhosis, liver cancer risk factors is very important. At present there are about 93 million of hepatitis B infection and HCV infection. 600~1000 host immune factors in hepatitis B and C the occurrence and development of disease susceptibility, and plays an important role in the response to treatment and so on. All play an important role in host innate immunity in NK cells, susceptibility and response to antiviral treatment and expression of single nucleotide polymorphism in chronic hepatitis C in patients with chronic hepatitis B of the occurrence and development of chemokine IP-10 in plasma. The implications of this experiment will explain the pathogenesis and host immune factors of efficacy of antiviral therapy of chronic hepatitis B and chronic hepatitis C.
Chemokines are chemotactic cytokines activity, plays an important role in inflammatory reaction. Many studies have confirmed that abnormal expression of hepatitis B virus disease of chemotactic factors closely related to the process. The high expression of chemokines can recruit a large number of effector cells such as cell toxicity to the liver in T lymphocytes remove the virus infected liver cells, but also because of the excessive release of chemokines, leading to inflammation expansion, exacerbated liver injury. Therefore clarify the mechanism of chemokines in viral hepatitis is particularly important.
IP-10 is found in 1985 of CXC chemokine subfamily members, plays an important role in the occurrence and development of liver immune injury and viral hepatitis. The studies show that high levels of IP-10 expression detection in patients with chronic hepatitis C in liver and plasma. Its high expression is closely related with the severity of inflammation and liver fibrosis. At present, the polymorphism of IP-10 and plasma science and outcomes and the relationship between the expression of response to interferon therapy after HCV infection is not clear.
NK cells play an important role in innate immune cells,.NK cells can directly kill virus infected cells in antiviral immunity and antibody mediated indirectly through the function of.NK cells play a cytotoxic effect on the cell mediated activation of receptor and inhibition with ligand cell surface receptors. The combination of these ligands include typical and atypical MHC class I antigen molecules, MHC like protein and various other autoimmune and virus derived. They may form or expression on the surface of infected cells.
There are few reports on the role and changes of NK cells in the process of HBV infection and the application of nucleotide analogues to antiviral therapy.
So the first part of this study by mass spectrometry and enzyme-linked immunosorbent assay technique: detection of patients with chronic hepatitis C, chronic hepatitis C virus infection after spontaneous clearance of the differential expression of plasma IP-10 in patients and healthy controls; observation of patients with chronic hepatitis C in interferon and ribavirin treatment, the expression changes of IP-10 in plasma in patients with antiviral the treatment and antiviral therapy; combined antiviral curative effect in the treatment of patients with chronic hepatitis C, found to predict the efficacy of the best IP-10 expression threshold; in patients with chronic hepatitis C, SNP locus rs3921 spontaneous clearance of IP-10 patients and healthy controls; rs8878; rs4859584; rs4241578; rs4859588; rs56061981; rs74810361; rs4256246 by sequencing analysis, analysis of different phenotypes in patients with chronic hepatitis C infection of different sites and IP-10 SNP The relationship between antiviral treatment and curative effect.
The first part of the experiment results are as follows: the spontaneous clearance of HCV plasma IP-10 expression than in patients with chronic hepatitis C and healthy controls were low, while healthy controls were lower than the expression in plasma of patients with chronic hepatitis C IP-10; for the treatment of patients with chronic hepatitis C interferon and Leigh Bhave Lin will induce the expression of plasma IP-10 increased, but 24 weeks after the end of treatment the plasma IP-10 the expression was significantly lower than before treatment; interferon combined with antiviral therapy in Leigh Bhave Lin before the baseline expression of IP-10 is less than 600pg/ml with easy access to complete early virological response independent predictive factors; interferon combined with antiviral therapy in Leigh Bhave Lin 2 weeks after the expression of IP-10 is less than 540pg/ml were the independent predictors to obtain early virological response to 8 SNP; in patients with chronic hepatitis C virus susceptibility loci detected in the IP-10 gene, and antiviral The curative effect was not related.
The second part of the study for the antiviral efficacy of tenofovir group and adefovir dipivoxil group with flow cytometry; the difference between the expression quantity and detection of NK cell receptor in patients with chronic hepatitis B in peripheral blood and normal controls; observe the tenofovir group and adefovir dipivoxil group changes in expression of NK cell receptor antiviral treatment process. And compare the differences between the two; find the NK cell receptor may predict the efficacy in patients with chronic hepatitis B virus.
The second part of the experimental results are as follows: for patients with hepatitis B and tenofovir, adefovir dipivoxil treatment with antiviral efficacy of different, tenofovir antiviral efficacy than adefovir dipivoxil; expression in patients with chronic hepatitis B and injury of NK cell receptor changes; expression of tenofovir or adefovir dipivoxil antiviral therapy can reduce NK cells in patients with chronic hepatitis B the inhibitory receptor NKG2A~+ and KIR2DL3~+; the expression of NKG2A~+ and KIR2DL3~+ receptor may be a predictor of curative effect evaluation of chronic hepatitis B patients.
In summary, IP-10 plays an important role in chronic hepatitis C infection and antiviral treatment, early treatment effect and the expression level of IP-10 can be directly predicted with the virus; NK cells play an important role in chronic hepatitis B infection, and the expression of NKG2A~ + and KIR2DL3~+ receptor may be a predictor of curative effect evaluation of chronic hepatitis B antiviral therapy in clinic.

【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2013
【分類號】：R512.62

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本文編號：1537617