摘要
乙型和丙型病毒性肝炎是两种呈世界性分布的传染性疾病,是急慢性肝病的主要原因。乙肝感染后会造成广泛的疾病谱,包括无症状的携带者,潜伏性肝炎、急性肝炎、慢性肝炎、爆发性肝炎,甚至肝硬化、原发性肝细胞癌。而丙型肝炎病毒(HCV)感染后,大约20%-30%感染者会发生自发性清除(Spontaneous Clearance),但70%-80%将会持续感染。在抗病毒治疗过程中,不同个体对干扰素治疗的应答有很大差异,约70%的病人经长效干扰素联合利巴韦林治疗一年后,可获得持久的病毒学应答(Sustained Virologic Response, SVR);而剩余30%患者对该治疗表现为无应答(No Response)。
这两种肝炎病毒感染后,疾病谱多样化,严重程度各有不同,对治疗的反应也大相径庭。大家普遍认为的病毒、环境、种族和患者的临床表现可能与疾病谱的多样化有关系,但尚不能完全解释疾病自然转归不同的真正原因。目前研究表明,宿主基因的变异是最重要的影响因素。尤其是近年来,国内外关于宿主遗传基因多态性与HCV感染转归的研究成果不断涌现,相继报道了诸如人类白细胞抗原、炎症趋化因子类,肿瘤坏死因子α(TNF-α)白介素和干扰素刺激基因等多个基因的多态性与HCV、HBV感染转归相关,都证实了宿主遗传基因变异可能导致机体免疫功能状态的差异,进而影响到HBV、HCV感染的不同临床结局。
IL28B基因编码IFN-λ3,为Ⅲ型干扰素位于19号染色体上,包含6个外显子,属于IFN-λs家族。近期在非洲、欧洲人群中进行的全基因组关联分析(Genome-wide Association Study, GWAS)鉴定发现编码IFNλ3的IL28B基因附近有一些单核苷酸多态位点与HCV病毒自发清除能力及对干扰素的应答有关,所以我们推测IL28B基因的变异及其表达可能与HCV、HBV的感染密切相关。
本文从宿主遗传易感性角度,借助国际最新关于慢性丙型肝炎宿主基因多态性的研究结果的启示,分别对丙肝感染者和乙肝感染者进行了IL28B rs12979860基因多态性及其表达水平的研究,同时对一部分丙肝患者进行了抗病毒治疗。收集我省汉族人口丙肝人群血样共896例(其中235例慢性丙肝患者参加抗病毒治疗),乙肝人群156例,所选人群符合Hardy-Weinberg平衡,通过焦磷酸测序法,分别对上述人群的IL28B SNP(rsl2979860)多态性位点(C/T)进行检测,应用ELISA试剂盒检测相关组个体血清中IL28B含量；应用Real Time PCR检测外周血单个核细胞(PBMC)中IL28B mRNA的表达情况。同时分析并评价不同基因型人群抗病毒治疗的应答情况。
结果发现,在丙肝人群研究中,中国健康人群中rs12979860 CC基因型频率为90.1%。自发清除组的C等位基因频率(rs12979860)高于于慢性感染组患者(P=0.04)。多变量Logistic回归分析,调整了性别和HBV共感染后,rs12979860基因多态性仍然是HCV自发清除的独立影响因素。在携带CC基因型患者中,HCV的自发清除率是T/C和T/T基因型患者的2倍(OR=2.13,P=0.04)。感染了HCV基因型1b型患者中,C/C基因型患者中HCV载量明显高于TC/TT基因型患者(P=0.014)。与C/C基因型患者相比,T/C或T/T基因型患者的转氨酶水平增高,差异显著。我们检测了HCV感染后不同临床转归的患者和健康人的IL28B血清水平以及其mRNA的表达水平,发现慢性HCV感染的患者中IL28B血清学和mRNA水平明显低于自发清除组和健康对照组人群。rs12979860 C/C基因型人群IL28B血清水平和IL28的mRNA表达水平都比携带T等位基因的人群稍高。在中国慢性丙肝患者的抗病毒治疗过程中,IL28B CC基因型的患者更可能获得病毒学应答,尤其是cEVR,多因素分析亦证实IL28B基因多态性是丙肝患者治疗成功与否的一项重要的预测因子。
在乙肝人群的研究中,我们共检测了156名HBV感染者和健康对照者,检测各组间IL28B血清学水平和IL28mRNA的表达水平,发现HBV携带组和健康对照组人群的IL28B无论是血清学还是mRNA的表达都明显高于其它各组(肝炎、肝硬化和肝癌组)(所有的P<0.05)。IL28B血清学水平在C/C基因型中的表达高于T/C基因型人群(P<0.05)。而在肝硬化+肝癌患者中,高转氨酶(ALT/AST)患者的IL28B无论在血清学还是mRNA水平都明显高于低转氨酶水平的患者(P<0.05)
总之,我们的结果表明,在中国人群中,IL28B上游的rs12979860 SNP及IL28B的表达不仅与HCV感染后的临床转归有关。而且也与HBV感染后的慢性化及疾病进展有关,同时也对慢性丙肝患者的抗病毒应答与否发挥重要作用。进一步研究也表明携带IL28B保护性等位基因能够引起IL28B表达水平的升高,IL28B在mRNA水平和血清学水平的表达两方面都证实,宿主IL28B基因多态性可能直接影响了IL28B的表达,而IL28B的表达水平及其功能变化则对HCV和HBV感染后的临床转归的进程中发挥重大作用,同时机体转氨酶水平增高,免疫应答反应增强的患者中,可能IL28B也积极的参与其中。可见,无论在HCV还是HBV感染的患者人群中,IL28B SNP位点变异及其表达水平改变可能是控制HCV、HBV复制的有益因素,IL28B基因多态性及其表达与疾病进展过程密不可分。
Hepatitis B and C are two infectious diseases in worldwide,which are the main reason of acute and chronic liver disease. HBV infection can cause a wide range of disease spectrum, including asymptomatic carriers, latent hepatitis, acute hepatitis, chronic hepatitis, fulminant hepatitis, and even cirrhosis, primary hepatocellular carcinoma. The hepatitis C virus (HCV) infection, approximately 20% -30% will clear the virus through spontaneous clearance, but 70% -80% will be persistent infection. In the course of antiviral treatment, the response to interferon therapy of HCV infection patients are very different, after Peg-interferon and ribavirin treatment, about 70% of the patients receive sustained virological response (SVR); while the remaining 30% of the patients showed no response to the treatment (No Response).
After infected by both hepatitis viruses, there are diversity, different severity of disease, and different response to therapy. We have a common view that the virus, environment, race and clinical manifestations of patients may be related with diversity of disease, but it can not fully explain the real reason of the natural outcome of the disease. Present study shows that the host gene mutation is the most important factor. Especially in recent years, the results of research on host genetic polymorphisms and outcome of HCV infection continue to emerge, which have been reported, such as human leukocyte antigen, inflammatory chemokines, tumor necrosis factorα(TNF-α),interleukin and interferon stimulated genes,that gene polymorphisms related to HCV, HBV infection outcomes.It have confirmed the host genetic variation may lead to differences in the body immune function, further influencing different clinical outcomes of HBV, HCV infection.
IL28B gene located on chromosome 19 interferon, consists of 6 exons, encoding IFN-λ3, as typeⅢ,are IFN-λs family. Recently Genome-wide Association Study(GWAS) of Africa, Europe population show that some single nucleotide polymorphisms near the IL28B coding IFNλ3 is related with spontaneous clearance capabilities of HCV viral and Response to interferon, so we speculated that the mutation and expression of IL28B gene may be associated closely with HCV, HBV infection.
From the perspective of host genetic susceptibility, with the enlightenment from the latest international host of chronic hepatitis C gene polymorphism results, We studied the L28B rs12979860 polymorphism and its expression level in HBV and HCV patients respectively, meanwhile, some chronic HCV infection patients accepted antviral therapy. We collected blood samples of Jilin province among the Han population, including 896 subjects of hepatitis C (among of them,235 chronic HCV infection patients accepted treatment),156 subjects of hepatitis B.the selected population accorded with Hardy-Weinberg equilibrium, these IL28B SNP (rs12979860) polymorphism (C/T) were detected respectively through the pyrosequencing method. ELISA test was used for IL28B serum concentration; the Real Time PCR was applicated for mRNA expression of IL28 of peripheral blood mononuclear cells (PBMC). At the same time, we also analysised the response status of the 235 HCV infection patients after treatment.
It was found that among populations of hepatitis C, the genotype frequency of rs12979860 CC of Chinese healthy population was 90.1%. C allele frequency (rs12979860) in spontaneous clearance group was higher than persistent infection patients (P=0.04). Maultivariate logistic regression analysis found, after adjusted gender and HBV infection, rs12979860 polymorphism is still an independent factor for HCV spontaneous clearance. Among patients carrying the C/C genotype, the rate of HCV spontaneous clearance are 2 times to T/C and T/T genotype patients (OR=2.13, P=0.04).
Patients infected HCV genotype lb, HCV load of patients with C/C genotype was significantly higher than T/C or T/T genotype (P=0.014). Comparing with C/C genotype patients, the patients with T/C or T/T genotype had higher ALT level, and the difference was significant. We detected the serum and mRNA expression levels of IL28B and found that in the chronic HCV infection patients, serology and IL28B mRNA levels were significantly lower than the spontaneous group and the healthy control group. In IL28B rs12979860 C/C genotype group, serum levels and mRNA expression levels of IL28B were slightly higher than the population carrying T allele.In the course of IFN/RBV treatment for Chinese chronic HCV infection patients, the patients carrying C/C genotype of IL28B could produced higher virologic response rate, especially cEVR, and multivariate logistic regression analysis also confirmed that the IL28B polymorphism is an important predictor for chronic hepatitis C patients treatment.
In the population infected hepatitis B, we detected a total of 156 HBV infected patients and healthy controls, testing serum levels and mRNA expression levels of IL28B in each groups, which found serum IL28B or mRNA expression of both groups(HBV carrier and healthy groups) were significantly higher than other groups (hepatitis, cirrhosis and liver cancer groups) (all P<0.05). Serum levels of IL28B of C/C genotype was higher than T/C genotype subjects (P<0.05). In the cirrhosis + hepatocellular carcinoma (HCC) patients, serum or IL28B mRNA levels with high aminotransferase (ALT/AST) of patient are significantly higher than patients with low transaminase levels (P<0.05)
In summary, our research show that in the Chinese population, the rs12979860 SNP located IL28B upstream and the expression of IL28B is not only related with clinical outcomes of HCV infection and the response of antivirologic treatment, but also with the chronic HBV infection and disease progression. Further studies have shown that carrying the protective allele of IL28B can cause elevated expression levels of IL28B. both the mRNA level and serum levels of IL28B confirmed that host gene IL28B polymorphisms may directly affect its expression, whereas expression and function of IL28B may play a major role on the clinical outcome of the process in the HCV and HBV infection, In patients with high transaminase level and strong immune responses, IL28B may also actively participate in it. So we believe that in patients both with HCV or HBV infection of the Chinese population, IL28B SNP variations and the change of the expression level may be a beneficial factor to control HCV, HBV replication. Gene polymorphism of IL28B and its expression are inseparable from disease progression.
引文
[1].Lavanchy D, The global burden of hepatitis C. Liver Int,2009; 29 Suppl 1:74-81.
[2].McHutchison JG, Everson GT, Gordon SC, Jacobson IM, Sulkowski M, et al., Telaprevir with peginterferon and ribavirin for chronic HCV genotype 1 infection. N Engl J Med,2009; 360(18):1827-38.
[3].Giorgini A, Capsoni F, Podda M, Lleo A, Battezzati PM, et al., Treatment with PEG-interferon and ribavirin for chronic hepatitis C increases neutrophil and monocyte chemotaxis. Ann N Y Acad Sci,2009; 1173:847-57.
[4].Knodell RG, Ishak KG, Black WC, Chen TS, Craig R, et al., Formulation and application of a numerical scoring system for assessing histological activity in asymptomatic chronic active hepatitis. Hepatology,1981; 1(5):431-5.
[5].Alric L, Fort M, Izopet J, Vinel JP, Charlet JP, et al., Genes of the major histocompatibility complex class Ⅱ influence the outcome of hepatitis C virus infection. Gastroenterology,1997; 113(5):1675-81.
[6]. Cramp ME, Carucci P, Underhill J, Naoumov NV, Williams R, et al., Association between HLA class Ⅱ genotype and spontaneous clearance of hepatitis C viraemia. J Hepatol, 1998; 29(2):207-13.
[7].Minton EJ, Smillie D, Neal KR, Irving WL, Underwood JC, et al., Association between MHC class Ⅱ alleles and clearance of circulating hepatitis C virus. Members of the Trent Hepatitis C Virus Study Group. J Infect Dis,1998; 178(1):39-44.
[8].Thio CL, Thomas DL, Goedert JJ, Vlahov D, Nelson KE, et al., Racial differences in HLA class Ⅱ associations with hepatitis C virus outcomes. J Infect Dis,2001; 184(1):16-21.
[9].Yoshizawa K, Ota M, Saito S, Maruyama A, Yamaura T, et al., Long-term follow-up of hepatitis C virus infection:HLA class Ⅱ loci influences the natural history of the disease. Tissue Antigens,2003; 61(2):159-65.
[10].Mangia A, Gentile R, Cascavilla I, Margaglione M, Villani MR, et al., HLA class Ⅱ favors clearance of HCV infection and progression of the chronic liver damage. J Hepatol, 1999;30(6):984-9.
[11].Aikawa T, Kojima M, Onishi H, Tamura R, Fukuda S, et al., HLA DRB1 and DQB1 alleles and haplotypes influencing the progression of hepatitis C. J Med Virol,1996; 49(4):274-8.
[12].Kuzushita N, Hayashi N, Moribe T, Katayama K, Kanto T, et al., Influence of HLA haplotypes on the clinical courses of individuals infected with hepatitis C virus. Hepatology,1998; 27(1):240-4.
[13].Kuzushita N, Hayashi N, Katayama K, Hiramatsu N, Yasumaru M, et al., Increased frequency of HLA DR13 in hepatitis C virus carriers with persistently normal ALT levels. J Med Virol,1996; 48(1):1-7.
[14].Peano G, Menardi G, Ponzetto A, and Fenoglio LM, HLA-DR5 antigen. A genetic factor influencing the outcome of hepatitis C virus infection? Arch Intern Med,1994; 154(23):2733-6.
[15].Congia M, Clemente MG, Dessi C, Cucca F, Mazzoleni AP, et al., HLA class Ⅱ genes in chronic hepatitis C virus-infection and associated immunological disorders. Hepatology,1996; 24(6):1338-41.
[16].Higashi Y, Kamikawaji N, Suko H, and Ando M, Analysis of HLA alleles in Japanese patients with cirrhosis due to chronic hepatitis C. J Gastroenterol Hepatol,1996; 11(3):241-6.
[17].Hohler T, Gerken G, Notghi A, Knolle P, Lubjuhn R, et al., MHC class Ⅱ genes influence the susceptibility to chronic active hepatitis C. J Hepatol,1997; 27(2):259-64.
[18].Tibbs C, Donaldson P, Underhill J, Thomson L, Manabe K, et al., Evidence that the HLA DQA1*03 allele confers protection from chronic HCV-infection in Northern European Caucasoids. Hepatology,1996; 24(6):1342-5.
[19].Verdon R, Pol S, Landais P, Mattlinger B, Carnot F, et al., Absence of association between HLA antigens and chronicity of viral hepatitis in haemodialyzed patients. J Hepatol, 1994;21(3):388-93.
[20].Yenigun A and Durupinar B, Decreased frequency of the HLA-DRB1*11 allele in patients with chronic hepatitis C virus infection. J Virol,2002; 76(4):1787-9.
[21].Hue S, Cacoub P, Renou C, Halfon P, Thibault V, et al., Human leukocyte antigen class Ⅱ alleles may contribute to the severity of hepatitis C virus-related liver disease. J Infect Dis,2002; 186(1):106-9.
[22].Tillmann HL, Chen DF, Trautwein C, Kliem V, Grundey A, et al., Low frequency of HLA-DRB1*11 in hepatitis C virus induced end stage liver disease. Gut,2001; 48(5):714-8.
[23].Barrett S, Ryan E, and Crowe J, Association of the HLA-DRB1*01 allele with spontaneous viral clearance in an Irish cohort infected with hepatitis C virus via contaminated anti-D immunoglobulin. J Hepatol,1999; 30(6):979-83.
[24].Fanning L, Kenny E, Sheehan M, Cannon B, Whelton M, et al., Viral load and clinicopathological features of chronic hepatitis C(1b) in a homogeneous patient population. Hepatology,1999; 29(3):904-7.
[25].Fanning LJ, Levis J, Kenny-Walsh E, Wynne F, Whelton M, et al., Viral clearance in hepatitis C(1b) infection:relationship with human leukocyte antigen class Ⅱ in a homogeneous population. Hepatology,2000; 31(6):1334-7.
[26].Kenny-Walsh E, Clinical outcomes after hepatitis C infection from contaminated anti-D immune globulin. Irish Hepatology Research Group. N Engl J Med,1999; 340(16):1228-33.
[27].Power JP, Lawlor E, Davidson F, Yap PL, Kenny-Walsh E, et al., Hepatitis C viraemia in recipients of Irish intravenous anti-D immunoglobulin. Lancet,1994; 344(8930):1166-7.
[28].Power JP, Lawlor E, Davidson F, Holmes EC, Yap PL, et al., Molecular epidemiology of an outbreak of infection with hepatitis C virus in recipients of anti-D immunoglobulin. Lancet,1995; 345(8959):1211-3.
[29].McKiernan SM, Hagan R, Curry M, McDonald GS, Nolan N, et al., The MHC is a major determinant of viral status, but not fibrotic stage, in individuals infected with hepatitis C. Gastroenterology,2000; 118(6):1124-30.
[30].Thio CL, Gao X, Goedert JJ, Vlahov D, Nelson KE, et al., HLA-Cw*04 and hepatitis C virus persistence. J Virol,2002; 76(10):4792-7.
[31].Lee WM, Hepatitis B virus infection. N Engl J Med,1997; 337(24):1733-45.
[32].Renou C, Halfon P, Pol S, Cacoub P, Jouve E, et al., Histological features and HLA class Ⅱ alleles in hepatitis C virus chronically infected patients with persistently normal alanine aminotransferase levels. Gut,2002; 51(4):585-90.
[33].Koziel MJ, Dudley D, Afdhal N, Grakoui A, Rice CM, et al., HLA class Ⅰ-restricted cytotoxic T lymphocytes specific for hepatitis C virus. Identification of multiple epitopes and characterization of patterns of cytokine release. J Clin Invest,1995; 96(5):2311-21.
[34].Hohler T, Kruger A, Gerken G, Schneider PM, Meyer zum Buschenefelde KH, et al., A tumor necrosis factor-alpha (TNF-alpha) promoter polymorphism is associated with chronic hepatitis B infection. Clin Exp Immunol,1998;111(3):579-82.
[35].Awad MR, El-Gamel A, Hasleton P, Turner DM, Sinnott PJ, et al., Genotypic variation in the transforming growth factor-beta1 gene:association with transforming growth factor-beta1 production, fibrotic lung disease, and graft fibrosis after lung transplantation. Transplantation,1998; 66(8):1014-20.
[36].Feder JN, Gnirke A, Thomas W, Tsuchihashi Z, Ruddy DA, et al., A novel MHC class Ⅰ-like gene is mutated in patients with hereditary haemochromatosis. Nat Genet,1996; 13(4):399-408.
[37].Hezode C, Cazeneuve C, Coue O, Pawlotsky JM, Zafrani ES, et al., Hemochromatosis Cys282Tyr mutation and liver iron overload in patients with chronic active hepatitis C. Hepatology,1998; 27(1):306.
[38].Smith BC, Gorve J, Guzail MA, Day CP, Daly AK, et al., Heterozygosity for hereditary hemochromatosis is associated with more fibrosis in chronic hepatitis C. Hepatology,1998; 27(6):1695-9.
[39].Wang SC, Ohata M, Schrum L, Rippe RA, and Tsukamoto H, Expression of interleukin-10 by in vitro and in vivo activated hepatic stellate cells. J Biol Chem,1998; 273(1):302-8.
[40].Poynard T, Bedossa P, and Opolon P, Natural history of liver fibrosis progression in patients with chronic hepatitis C. The OBSVIRC, METAVIR, CLINIVIR, and DOSVIRC groups. Lancet,1997; 349(9055):825-32.
[41].Powell EE, Edwards-Smith CJ, Hay JL, Clouston AD, Crawford DH, et al., Host genetic factors influence disease progression in chronic hepatitis C. Hepatology,2000; 31(4):828-33.
[42].Bedossa P and Paradis V, Transforming growth factor-beta (TGF-beta):a key-role in liver fibrogenesis. J Hepatol,1995; 22(2 Suppl):37-42.
[43].Gibbons GH, Pratt RE, and Dzau VJ, Vascular smooth muscle cell hypertrophy vs. hyperplasia. Autocrine transforming growth factor-beta 1 expression determines growth response to angiotensin Ⅱ. J Clin Invest,1992; 90(2):456-61.
[44].Marcellin P, Hepatitis B and hepatitis C in 2009. Liver Int,2009; 29 Suppl 1:1-8.
[45].Seeff LB, Natural history of chronic hepatitis C. Hepatology,2002; 36(5 Suppl 1):S35-46.
[46].Shiffman ML, Treatment of hepatitis C in 2011:what can we expect? Curr Gastroenterol Rep,2010; 12(1):70-5.
[47].Muller R, The natural history of hepatitis C:clinical experiences. J Hepatol,1996; 24(2 Suppl):52-4.
[48].Sheppard P, Kindsvogel W, Xu W, Henderson K, Schlutsmeyer S, et al., IL-28, IL-29 and their class Ⅱ cytokine receptor IL-28R. Nat Immunol,2003; 4(1):63-8.
[49].Kotenko SV, Gallagher G, Baurin VV, Lewis-Antes A, Shen M, et al., IFN-lambdas mediate antiviral protection through a distinct class II cytokine receptor complex. Nat Immunol,2003; 4(1):69-77.
[50].Sommereyns C, Paul S, Staeheli P, and Michiels T, IFN-lambda (IFN-lambda) is expressed in a tissue-dependent fashion and primarily acts on epithelial cells in vivo. PLoS Pathog,2008; 4(3):e1000017.
[51].Ge D, Fellay J, Thompson AJ, Simon JS, Shianna KV, et al., Genetic variation in IL28B predicts hepatitis C treatment-induced viral clearance. Nature,2009; 461(7262):399-401.
[52].Kau A, Vermehren J, and Sarrazin C, Treatment predictors of a sustained virologic response in hepatitis B and C. J Hepatol,2008; 49(4):634-51.
[53].王东生,潘煜,牛俊奇,姜晶,姜艳芳,等.,IL-28B基因变异与丙型肝炎的易感性及其自然清除的关系.临床肝胆病杂志,2010；26(5)：496-499.
[54].McCarthy JJ, Li JH, Thompson A, Suchindran S, Lao XQ, et al., Replicated association between an IL28B gene variant and a sustained response to pegylated interferon and ribavirin. Gastroenterology,2010; 138(7):2307-14.
[55].Montes-Cano MA, Garcia-Lozano JR, Abad-Molina C, Romero-Gomez M, Barroso N, et al., Interleukin-28B genetic variants and hepatitis virus infection by different viral genotypes. Hepatology,2010; 52(1):33-7.
[56].Tanaka Y, Nishida N, Sugiyama M, Kurosaki M, Matsuura K, et al., Genome-wide association of IL28B with response to pegylated interferon-alpha and ribavirin therapy for chronic hepatitis C. Nat Genet,2009; 41(10):1105-9.
[57].Suppiah V, Moldovan M, Ahlenstiel G, Berg T, Weltman M, et al., IL28B is associated with response to chronic hepatitis C interferon-alpha and ribavirin therapy. Nat Genet,2009; 41(10):1100-4.
[58].Rauch A, Kutalik Z, Descombes P, Cai T, Di Iulio J, et al., Genetic variation in IL28B is associated with chronic hepatitis C and treatment failure:a genome-wide association study. Gastroenterology,2010; 138(4):1338-45,1345 e1-7.
[59].Thomas DL, Thio CL, Martin MP, Qi Y, Ge D, et al., Genetic variation in IL28B and spontaneous clearance of hepatitis C virus. Nature,2009; 461(7265):798-801.
[60].Grebely J, Petoumenos K, Hellard M, Matthews GV, Suppiah V, et al., Potential role for interleukin-28B genotype in treatment decision-making in recent hepatitis C virus infection. Hepatology,2010; 52(4):1216-24.
[61].Rauch A, Rohrbach J, and Bochud PY, The recent breakthroughs in the understanding of host genomics in hepatitis C. Eur J Clin Invest,2010; 40(10):950-9.
[62].Sarasin-Filipowicz M, Oakeley EJ, Duong FH, Christen V, Terracciano L, et al., Interferon signaling and treatment outcome in chronic hepatitis C. Proc Natl Acad Sci U S A, 2008; 105(19):7034-9.
[63].Urban TJ, Thompson AJ, Bradrick SS, Fellay J, Schuppan D, et al., IL28B genotype is associated with differential expression of intrahepatic interferon-stimulated genes in patients with chronic hepatitis C. Hepatology,2010; 52(6):1888-96.
[64].于淑丽,龚幼龙和邵瑞太.,慢性乙肝、乙肝后肝硬化和肝癌的疾病负担.中国公共卫生,2003；19(3)：280-282.
[65].Thursz M, Genetic susceptibility in chronic viral hepatitis. Antiviral Res,2001; 52(113-116).
[66].Amundadottir L, Thorvaldsson S, Gudbjartsson D, and etal., Cancer as a complex phenotype:pattern of cancer distribution within and beyond the nuclear family. PloS Med, 2004; 1:e65.
[67].Karayiannis P, Alexopoulou A, Hadziyannis S, Thursz M, and etal., Fulminant hepatitis associated with hepatitis B virus e antigen-negative infection:importance of host factors. Hepatology,1995; 22:1628-1643.
[68].Weatherall D, Clegg J, and Kwiatkowski D, The role of genomics in studying genetic susceptibility to infectious disease. Genome Res,1997; 7:967-973.
[69].de Araujo ES, Dahari H, Cotler SJ, Layden TJ, Neumann AU, et al., Pharmacodynamics of PEG-IFN alpha-2a and HCV response as a function of IL28B polymorphism in HIV/HCV co-infected patients. J Acquir Immune Defic Syndr,2010.
[70].Kao J-H, Chen P-j, Lai M--Y, and al. e, Hepatitis B genotypes corelate with clinical outcomes in patientst with chronic hepatitis B. Gastroenterology,2000; 118:554-559.
[71].Lin T, Chen C, Wu M, Yang C, Chen J, et al., Hepatitis B virus markers in Chinese twins. Anticancer Res,1989; 9:737-741.
[72].Amundadottir L, Thorvaldsson S, Gudbjartsson D, and etal., Cancer as a complex phenotype:pattern of can cer distribution within and beyond the nuclear family. PLoSled, 2004; 1:e65.
[73].Akbar N, Basuki B, Garabrant D, Sulaiman A, and Noer H, Ethnicity, socioeconomic status, transfusions and risk of hepatitis B and hepatitis C infection. J Gastroenterol Hepatol,1997; 12:752-757.
[74].Chin P, Chu D, Clarke K, Odom-Maryon T, Yen Y, et al., Ethnic differences in the behavior of hepatocellular carcinoma. Cancer,1999; 85:1931-1936.
[75].Zhao S, Li H, Lou H, and etal., High Prevalence of HBV in Tibet.China.Asian Pac. J Cancer Prev,2001; 2:299-304.
[76].Evans A, O'Connell A, Pugh J, Mason W, and Shen F, Geographic variation in viral load among hepatitis B carriers with differing risks of hepatocellular carcinoma. Cancer Epidemiol Biomarkers Prev,1998; 7:559-565.
[77].Miller L and Hendrie N, Health of children adopted from China. Pediatrics,2000; 105:e76.
[78].London W and Drew S, Sex differences in response to hepatitis B infection among patients receiving chronic dialysis treatment.. Proc Natl Acad Sci USA,1977; 74:2561-2653.
[79].Lee C, Lu S, Changchien C, Yeh C, Hsu T, et al., Age, gender and local geographic variations of viral etiology of hepatocellular carcinoma in a hyperendemic area of hepatitis B virus infection. Cancer,1999; 86:1143-1150.
[80].Bosch F and Ribes J, Epidemiology of liver cancer in Europe. Can J Gastroenterol, 2000; 14:621-630.
[81].Blakely T, Bates M, Garrett N, and Robson B, The incidence of hepatocellular carcinoma in New Zealand. N Z Med J,1998; 111:471-474.
[82].Realdi G, Fattovich G, Pastore G, Caredda F, and Noventa F, Problems in the management of chronic hepatitis B with interferon:experience in a randomized multicentre study. J Hepatol,1990; 11((Suppl 1)):s129-s132.
[83].Craxi A, Montano L, Goodall A, and Thomas H, Genetic and sex-linked factors influencing HBs antigen clearance,I,Nonimmune clearance in inbred strains of mice. J Med Virol,1982; 9:117-123.
[84].白汉玉.和张春芳.,小儿病毒性肝炎的流行病学、临床表现、诊断、预后及治疗研究.病毒性肝炎防治研究,1993：p201-207.
[85].Mulligen M and Stiehm E, Neonatal hepatitis B infection:clinical and immunologic considerations. J Perinatol,1994; 14:2-9.
[86].Ryder R. Whittle H. Wojiecowsky T, and Moffat W, Screening for hepatitis B virus markers is not justified in West African transfusion centres. Lancet,1984; 2:449-452.
[87].Thursz M, Kwiatkowski D, Allsopp C, Greenwood B, and Thomas H, Association between an MHC class Ⅱ allele and clearance of hepatitis B virus in The Gambia. N Engl J Med,1995; 332:1065-1069.
[88].Lok A, Lai C, Chung H, Lau J, Leung E, et al., Morbidity and mortality from chronic hepatitis B virus infection in family members of patients with malignant and nonmalignant hepatitis B virus-related chronic liver disease. Hepatology,1991; 13:834-837.
[89].Knolle P, Kremp S, Hohler T, Krummenauer F, and Schirmacher P, Viral and host factors in the prediction of response to interferon-alpha therapy in chronic hepatitis C after long-term follow-up. J Viral Hepat 1998; 5:399-406.
[90].Thio C, Thomas D, and Carrington M, Chronic viral hepatitis and the human genome. Hepatology 2000; 31:819-827.
[91].Sachidanandam R, Weissman D, and Schmidt S, A map of human genome sequence variation containing 1.42 million single nucleotide polymorphisms. Nature,2001; 409:928-933.
[92].Matise T, Sachidanandam R, and Clark A, A 3.9-centimorgan -resolution human single nucleotide polymorphism linkage map and screening set. Am J Human Genet,2003; 73:271-284.
[93].Dean M, Carrington M, and Wialder C, Genetic restriction of HIV-1 infection and progression to AIDS by a deletion allele of the CKR5 structural gene.Hemophilia Growth and Development Study,multicenter AIDS Cohort Study,Multicenter Hemophilia Cohort Study,San Francisco City Cohort,ALIVE Study. Science,1996; 273:1856-1862.
[94].Dean M, Carrington M, and O'Brien S, Balanced polymorphism selected by genetic versus infectious human disease. Annu Rev Genomics Hum Genet 2002; 3:263-292.
[95].Smith C, Genomics:SNPs and human disease. Nature,2005; 435:993.
[96].Thursz M, Pros and cons of genetic association studies in hepatitis B. Hepatology, 2004; 40:284-286.
[97].王宇明和顾长海.,感染病学新进展.北京：人民卫生出版社,2001；第一版：216-233.
[98].Chloe L T, David L T, Peter K, and al. e, Comprehensive Analysis of Class I and Class II HLA Antigens and Chronic Hepatitis B Virus Infection. Journal of Virology,2003; 77:12083-12087.
[99].Thursz M, Genetic susceptibility in chronic viral hepatitis. Antiviral Res,2001; 52:113-116.
[100].Thursz M, Kwiatkowski D, Allsopp C, Greenwood B, Thomas H, et al., Association between an MHC class Ⅱ allele and clearance of hepatitis B virus in the Gambia. N Engl J Med 1995; 332:1065-1069.
[101].Hohler T, Gerken G, Notghi A, Lubjuhn R, Taheri H, et al., HLA-DRB1*1301 and*1302 protect against chronic hepatitis B. J Hepatol,1997; 26:503-507.
[102].Diepolder H, Jung M, Keller E, Schraut W, Gerlach J, et al., A vigorous virus-specific CD4+ T cell response may contribute to the association of HLA-DR13 with viral clearance in hepatitis B.. Clin Exo Immunol 1998; 113:244-251.
[103].Westendorp R, Langermans J, and Huizinga T, Genetic influence on cytokine production in meningococcal disease. Lancet 1997; 349:1912-1913.
[104].Van Devanter S, Cytokine and cytokine receptor polymorphisms in infectious disease. Intensive Care Med 2000; 26((Suppl 1)):S98-102.
[105].Knight J and Kwiatkowski D, Inherited variability of tumor necrosis factor production and susceptibility to infectious disease. Pro Assoc Am Physicians,1999; 111:290-298.
[106].Juszczynski P, Kalinka E, Bienven J, and etal., Human leukocyte antigens class II and tumor necrosis factor genetic polymorphisms are independent predictors of non-Hodgkin lymphoma outcome. Blood 2002; 100:3037-3040.
[107].Higuchi T, Seki N, Kamizono S, Yamada A, and etal., Polymorphism of the 5'-flanking region of the human tumor necrosis factor(TNF)-alpha gene in Japanese. Tissue Antigens,1998; 51:605-612.
[108].Hohler T, Kruger A, Gerken G, Schneider P, and etal., A tumor necrosis factor-alpha (TNF-alpha)promoter polymorphism is associated with chronic hepatitis B infection.. Clin Exp Immunol 1998; 111:579-582.
[109].Hohler T, Kruger A, Gerken G, Schneider P, and etal., Tumor necrosis factor-alpha promoter polymorphism at position-238 is associated with chronic hepatitis B infection. J Med Virol 1998; 54:173-177.
[110].Wilson A, De Vries N, Pociot F, and etal., An allelic polymorphism within the human tumor necrosis factor alpha promoter region is strongly associated with HLA A1, B8, and DR3 alleles. J Exp Med,1993; 177:557-560.
[111].D'Alfonso S and Richiardi P, A polymorphic variation in a putative regulation box of the TNFA promoter region. Immunogenetics,1994; 39:150-154.
[112].Yee L, Tang J, Herrera J, and al. e, Tumor necrosis factor gene polymorphisms in patients with cirrhosis from chronic hepatitis C virus infection. Genes Immun,2000; 1(386-390).
[113].Rosen H, McHutchison J, Conrad A, and al. e, Tumor necrosis factor genetic polymorphismsand response to antiviral therapy in patients with chronic hepatitis C. Am J Gastroenterol,2002; 97:714-720.
[114].Schiemann U, Glas J, Torok P, and al. e, Response to combination therapy with interferon alfa-2a and ribavirin in chronic hepatitis C according to a TNF-alpha promoter polymorphism. Digestion,2003; 68:1-4.
[115].Yu M, Dai C, Chiu C, and al. e, Tumor necrosis factor alpha promoter polymorphisms at position-308 in Taiwanese chronic hepatitis C patients treated with interferon-alpha. Antiviral Res,2003; 59:35-40.
[116].Miyazoe S, Hamasaki K, Nakata K, and etal., Influence of interleukin-10 gene promoter polymorphisms on disease progression in patients chronically infected with hepatitis B virus.. Am J Gastroenterol 2002; 97:2086-2092.
[117].Ben-Ari Z, Mor E, Papo O, and etal., Cytokine gene polymorphisms in patients infected with hepatitis B virus. Am J Gastroenterol,2003; 98:144-150.
[118].顾绍庆,朱启镕,俞蕙,等.,肿瘤坏死因子-α基因多态性与乙型肝炎病毒宫内感染易感性的关系.中华肝脏病杂志,2004；12：538-539.
[119].Tokushige K, Tsuchiya N, Hasegawa K,., and al. e, Influence of TNF gene polymorphism andHLA-DRB1 haplotype in Japanese patients with chronic liver disease caused by HCV. Am J Gastroenterol 2003; 98:160-166.
[120].Kim Y, Lee H, Yoon J, and al. e, Association of TNF-a promoter polymorphismswith the clearance of hepatitis B virus infection. Hum Mol Gene,2003; 12:2541-2546.
[121].Thio C, Goedert J, Mosbruger T, and al. e, An analysis of tumor necrosis factor alpha gene polymorphisms and haplotypes with natural clearance of hepatitis C virus infection. Genes Immun,2004; 5:294-300.
[122].Chevillard C, Henri S, Stefani F, and al. e, Two new polymorphisms in the human interferon gamma(IFN-gamma)promoter. Eur J Immuno genet,2002; 29:53-56.
[123].Pravica V, Perrey C, Stevens A, and al. e, A single nucleotide polymorphism in the first intron of the human IFN-gamma gene:absolute correlation with a polymorphic CA microsatellite marker of high IFN-gamma production. Hum Immunol,2000; 61:863-866.
[124].Henri S, Stefani F, Parzy D, and et al., Description of three new polymorphisms in the intronic and 3'UTR regions of the human interferon gamma gene. Genes Immun,2002; 3:1-4.
[125].Hoffmann S, Stanley E, Cox E, and etal., Ethnicity greatly influence cytokine gene polymorphism distribution. Am J Transplant 2002; 2:560-567.
[126].Ben-Ari Z, Mor E, Papo O, and etal., Cytokine gene polymorphisms in patients infected with hepatitis B virus. Am J Gastroenterol 2003; 98:144-150.
[127].Redpath S, Ghazal P, and Gascoigne N, Hijacking and exploitation of IL-10 by intracellular pathogens. Trends Microbial,2001; 9:86-92.
[128].Asadullah K, Sterry W, and Volk H, Interleukin-10 Therapy-Review of a New Approach. Pharmacol.Rev 2003; 55:241-269.
[129].Edwards-Smith C, Jonsson J, and etal., Interleukin-10 promoter polymorphism predicts initial response of chronic hepatitis C to interferon alfa. Hepatology,1999; 30:526-530.
[130].Mangia A, Santoro R, Piattelli M, and et al., IL-10 haplotypes as possible predictors of spontaneous clearance of HCV infection. cytokine 2004; 25:103-109.
[131].Miyazoe S, Hamasaki K, Nakata K, Kajiya Y, and. et al, Influence of interleukin-10 gene promoter polymorphisms on disease progression in patients chronically infected with hepatitis B virus. Am J Gastroenterol 2002; 97:2086-2092.
[132].Yee L, Tang J, Gibson A, Kimberly R, and et al. Interleukin 10 polymorphisms as predictors of sustained response in antiviral therapy for chronic hepatitis C infection. Hepatology,2001; 33:708-712.
[133].Rossi D and Zlotnik A, The biology of chemokines and their receptors. Annu Rev Immunol,2000; 18:217-242.
[134].Narumi S, Tominaga Y, Tamaru M, and et al., Expression of IFN-inducible protein-10 in chronic hepatitis.. J Immuno,1997; 158:5536-5544.
[135].Samson M, Libert F, Doranz B, and e tal., Resistance to HIV-1 infection in Caucasian individuals bearing mutant alleles of the CCR5 chemokine receptor gene. Nature, 1996; 382:722-725.
[136].Woitas RP, Ahlenstiel G, Iwan A, and etal., Frequency of the HIV-protective CC chemokine receptor 5-Delta32/Delta32 genotype is increased in hepatitis C. Gastroenterology 2002; 122:1721-1728.
[137].Promrat K, McDermott D, Gonzalez C, and etal., Associations of chemokine system polymorphisms with clinical outcomes and treatment responses of chronic hepatitis C.. Gastroenterology,2003; 124:352-360.
[138].李定国和刘清华.,雌激素与肝纤维化研究现状.国外医学.消化系疾病分册.,2005；24(4)：195-197.
[139].荣顺兴,金泰澳.,.雌激素受体的研究进展.职业卫生与应急救援.,2005；23(1)：11-14.
[140].Mizoguchi Y, Takeda H, Kobayashi K, and etal., Impairment in the response of peripheral blood mononuclear cells from asymptomatic hepatitis B virus carriers to estradiol.. Jpn J Med 1988; 27:183-186.
[141].Mizoguchi Y, Takeda H, Sakagami Y, and etal., Estradiol receptors in the cytosol of peripheral blood mononuclear cells in hepatitis B virus carriers treated with interferon-alpha.. Gastroenterol Jpn,1989; 24:373-379.
[142].Vilia E, Camellini L, Dogani A, and etal., Variant estrogen receptors mNA species detected in human primary hepatocellular carcinoma. Caneer Res,1995; 55:498-500.
[143].Smith E, Boyd J, Frank G, and etal., Estrogen resistance caused by a mutation in the estrogen-receptor gene in a man. N Engl J Med 1994; 331:1056-1061.
[144].Deng G, Zhou G, Zhai Y, and etal., Association of estrogen receptor a polymorphisms with susceptibility to chronic hepatitis B virus infection. Hepatology 2004; 40:318-326.
[145].Kuhlman M, Joiner K, and Ezekowitz R, The human mannose binding protein functions as an opsonin.. J Exp Med,1989; 169:1733-1745.
[146].Lipscombe R, Sumiya M, Hill A, and etal., High frequency in African and non-African populations of independent mutations in the mannose binding protein gene. Hum Mol Genetic,1992; 1:709-715.
[147].Thomas H, Foster G, Sumiya M, and etal., Mutation of gene of mannose-binding protein associated with chronic hepatitis B viral infection. Lancet,1996; 348:1417-1419.
[148].Hohler T, Wunschel M, Gerken G, and etal., No association between mannose binding lectin alleles and susceptibility to chronic hepatitis B virus infection in German patients.. Exp Clin Immunogenet,1998; 15:130-133.
[149].Dai Y, Ning T, Li K, Qi SX, and etal., Association between LMP2/LMP7 gene polymorphism and the infection of hepatitis B virus.. Beijing Da Xue Xue Bao,2005; 37:508-512.
[150].Bellamy R, Ruwende C, Corrah T, and etal., Tuberculosis and chronic hepatitis B virus infection in Africans and variation in the vitamin D receptor gene.. J Infect Dis 1999; 179:721-724.
[151].单晶,王璐,李卓等.,维生素D受体基因多态性与乙型肝炎阳性者家庭聚集性关系.中国医学科学院学报2006；28：148-153.
[152].李俊红,陈冬梅,李卓等.,维生素D受体基因多态性与乙型肝炎病毒感染的关联研究..中华医学遗传学杂志,2006；23：402-405.
[153].Mohammad Alizadeh A, Hajilooi M, Ranjbar M, and etal., Cytotoxic T-lymphocyte antigen 4 gene polymorphisms and susceptibility to chronic hepatitis B. World J Gastroenterol,2006; 12:630-635.
[154].陈涛,陈明,汪莉萍,封波,李秀华.,CD28基因启动子-372G/A多态性与慢性乙型肝炎的关系..中华肝脏病杂志,2006；14：312-313.
[155].McMahon B, The natural history of chronic hepatitis B virus infection.. Semin Liver Dis 2004; 24(Suppl 1):17-21.
[156].Abel L and Dwssein A, Genetic epidemiology of infectious diseases in humans:design of population-based studies.. Emerge Infect Dis,1998; 4:593-603.
[157].Griffiths P, Interactions between viral and human genes.. Rev Med Virol 2002; 12:197-199.
[158].Ank N, Iversen MB, Bartholdy C, Staeheli P, Hartmann R, et al., An important role for type Ⅲ interferon (IFN-lambda/IL-28) in TLR-induced antiviral activity. J Immunol, 2008; 180(4):2474-85.
[159].姚光弼.,规范乙型肝炎防治行为的重要性.中华传染病杂志,2007；25(1)：59-60.
[160].Aparicio E, Parera M, Franco S, Perez-Alvarez N, Tural C, et al., IL28B SNP rs8099917 is strongly associated with pegylated interferon-alpha and ribavirin therapy treatment failure in HCV/HIV-1 coinfected patients. PLoS One,2010; 5(10):e13771.
[161].Li JH, Lao XQ, Tillmann HL, Rowell J, Patel K, et al., Interferon-lambda genotype and low serum low-density lipoprotein cholesterol levels in patients with chronic hepatitis C infection. Hepatology,2010; 51(6):1904-11.
[162].Doyle SE, Schreckhise H, Khuu-Duong K, Henderson K, Rosier R, et al., Interleukin-29 uses a type 1 interferon-like program to promote antiviral responses in human hepatocytes. Hepatology,2006; 44(4):896-906.
[163].Langhans B, Kupfer B, Braunschweiger I, Arndt S, Schulte W, et al., Interferon-lambda serum levels in hepatitis C. J Hepatol,2010.
[164].Ghany MG, Strader DB, Thomas DL, and Seeff LB, Diagnosis, management, and treatment of hepatitis C:an update. Hepatology,2009; 49(4):1335-74.
[165].Huang CF, Yang JF, Huang JF, Dai CY, Chiu CF, et al., Early identification of achieving a sustained virological response in chronic hepatitis C patients without a rapid virological response. J Gastroenterol Hepatol,2010; 25(4):758-65.
[166].Davis GL, Wong JB, McHutchison JG, Manns MP, Harvey J, et al., Early virologic response to treatment with peginterferon alfa-2b plus ribavirin in patients with chronic hepatitis C. Hepatology,2003; 38(3):645-52.
[167].Yu ML, Huang CF, Huang JF, Chang NC, Yang JF, et al., Role of interleukin-28B polymorphisms in the treatment of hepatitis C virus genotype 2 infection in Asian patients. Hepatology,2011; 53(1):7-13.
[168].Lu HY, Zhuang LW, Yu YY, Si CW, Li J, et al., Effects of antiviral agents and HBV genotypes on intrahepatic covalently closed circular DNA in HBeAg-positive chronic hepatitis B patients. World J Gastroenterol,2008; 14(8):1268-73.
[169].Preiss S, Littlejohn M, Angus P, Thompson A, Desmond P, et al., Defective hepatitis B virus DNA is not associated with disease status but is reduced by polymerase mutations associated with drug resistance. Hepatology,2008; 48(3):741-9.
[170].李明才,何韶衡.,重组人IL-29的原核表达、纯化及活性分析.第四军医大学学报,2006；23.
[171].O'Brien TR, Interferon-alfa, interferon-lambda and hepatitis C. Nat Genet,2009; 41(10):1048-50.
[172].中华医学会肝病学分会and中华医学会感染病学分会.,慢性乙型肝炎防治指南.现代消化及介入治疗,2006；11(4)：245-255.
[173].中国抗癌协会肝癌专业委员会.,原发性肝癌诊断标准.中华肝脏病杂志,2000;8：135-135.
[174].Thursz M, Genetic susceptibility in chronic viral hepatitis. Antiviral Res,2001; 52(2):113-6.
[175].Auffermann-Gretzinger S, Keeffe EB, and Levy S, Impaired dendritic cell maturation in patients with chronic, but not resolved, hepatitis C virus infection. Blood,2001; 97(10):3171-6.
[176].Beckebaum S, Cicinnati VR, Zhang X, Ferencik S, Frilling A, et al., Hepatitis B virus-induced defect of monocyte-derived dendritic cells leads to impaired T helper type 1 response in vitro:mechanisms for viral immune escape. Immunology,2003; 109(4):487-95.