Autophagy-Related 5 Gene rs510432 Polymorphism Is Associated with Hepatocellular Carcinoma in Patients with Chronic Hepatitis B Virus Infection

Background: Despite the identification of autophagy-related protein 5 (ATG5) as a molecule involved in the activated autophagy machinery during hepatitis B virus (HBV) infection and hepatocarcinogenesis, the consequences of ATG5 mutation carriage for patients with chronic HBV infection remain unclear. This study examined the association of ATG5 polymorphisms with HBV-related diseases including hepatocellular carcinoma (HCC).

Patients and Methods: Two functionally relevant polymorphisms ATG5 rs573775 and rs510432 were genotyped by ligase detection reaction-polymerase chain reaction in 403 patients with chronic HBV infection (171 chronic hepatitis, 119 cirrhosis and 113 HCC) and 196 healthy controls. Univariate and multivariate logistic regression was performed to evaluate factors associated with HCC.

Results: The rs573775 genotype and allele frequencies had no significant differences between patients with different clinical diseases. However, HCC patients had significantly higher frequency of rs510432 genotype AA (odds ratio [OR] 2.185, 95% confidence interval [CI] 1.042–4.581, P = 0.037, P value by Bonferroni correction [P_c] = 0.074) and allele A (OR 1.435, 95% CI 1.023–2.013, P_c = 0.036) than chronic hepatitis patients. In multivariate analyses, rs510432 allele A-containing genotypes (AA+GA) were independently associated with cirrhosis in comparison to chronic hepatitis (OR 1.927, 95%CI 1.011–3.017, P = 0.032). The rs510432 genotypes AA+GA were also independently associated with HCC in comparison to chronic hepatitis (OR 2.583, 95% CI 1.025–3.911, P = 0.006) or chronic HBV infection without HCC (OR 2.632, 95% CI 1.067–3.482, P = 0.032).

Conclusion: These results indicate that rs510432 genotypes AA+GA are associated with disease progression and HCC risk in chronic HBV infection, providing novel evidence for a role of ATG5 in the pathogenesis of HBV-related HCC.

Abbreviations: HBV: hepatitis B virus; HCC hepatocellular carcinoma; TNFSF10: tumor necrosis factor superfamily member 10; ATG5: autophagy-related protein 5; DNA: deoxyribonucleic acid; LDR-PCR: ligase detection reactions-polymerase chain reaction; PCR: polymerase chain reaction; SLE: systemic lupus erythematosus; BD: Behçet’s disease; IL-10: interlukin-10; LPS: lipopolysaccharide; PBMC: peripheral blood mononuclear cells; CWP: coal workers’ pneumoconiosis; TNF-α: tumor necrosis factor-α     

Hepatitis C virus infection among dialysis patients in Tunisia: incidence and molecular evidence for nosocomial transmission

In order to study the incidence of hepatitis C virus (HCV) infection in Tunisian haemodialysis patients and detect its nosocomial transmission, 395 patients were enrolled in a prospective study (November 2001-2003). HCV serological and virological status was determined initially using, respectively a third generation ELISA and an RT-PCR qualitative assay. The genotype of the HCV isolates was determined by sequencing NS5B region. The issue of nosocomial transmission was addressed by sequencing the HVR-1 region of the E2 gene. About 20% of the patients had anti-HCV antibodies and HCV-RNA was detected in 73% of the anti-HCV positive patients. Two cases of de novo HCV infection were identified in two dialysis centers, during virological follow-up of patients susceptible to HCV infection. The incidence of de novo HCV infection was 0.5%. Determining the genotypes in the first center disclosed that all HCV-positive patients were infected with genotype 1b; sequencing of the HVR-1 region of the E2 gene provided strong evidence that the isolate from the newly infected patient and another infected dialysis patient were closely related, confirming nosocomial contamination. The investigation of the second center is pending. Besides, one patient with negative HCV serology had detectable HCV-RNA at the beginning of the study. This case had HCV genotype 1b, two other infected dialysis patients in the same unit had HCV genotypes 4k and 3a; thus precluding nosocomial transmission. Thanks to molecular and phylogenetic methods, one case of nosocomial HCV transmission in haemodialysis was confirmed. Epidemiological investigation suggested nosocomial transmission via the medical and/or nursing staff.     

Association of genetic variations in CCR5 and its ligand, RANTES with clearance of hepatitis B virus in Korea

Immunogenetic factors may play a role in determining the susceptibility of an individual to viral infection. The aim of current study was to investigate the association of clearance of hepatitis B virus (HBV) with promoter polymorphisms within the CC chemokine receptor 5 (CCR5) and its major ligand, regulated upon activation, normal T cells expressed and secreted (RANTES) genes. Five chemokine system polymorphisms (CCR5 Delta32, CCR5 promoter 59029G/A, 59353C/T, RANTES -403G/A, and -28C/G) were studied in a total of 698 subjects. The carriage of each genetic variant was compared among "spontaneously recovered" group (n = 243), "chronic carrier" group (n = 349), and "unexposed" group (n = 106). CCR5 59029G promoter variant was associated with clearance of HBV infection in an acute phase (OR = 1.71, P = 0.006, dominant model; OR = 2.17, P < 0.001, recessive model) and amelioration of hepatic inflammation (P = 0.003) with the control of HBV replication (P = 0.04) in chronic carriers. Interestingly, CCR5 59029 was linked completely to CCR5 59353, and CCR5 Delta32 homozygosity or heterozygosity was not found in any Korean patient. No association was seen with RANTES polymorphisms at position -403 and -28. The CCR5 59029G/CCR5 59353T polymorphism may play a role in the clearance of HBV infection.     

A polymorphism in melanoma differentiation-associated gene 5 may be a risk factor for enterovirus 71 infection

Enterovirus 71 (EV71) infection has a wide variety of clinical manifestations, from no symptoms to fatal disease. Host immune response may be a determinant of disease severity. We investigated the association of polymorphisms in three pattern recognition receptor (PRR) genes—toll-like receptor 3 (TLR3) (rs3775291), retinoic acid-inducible gene I (RIG-I) (rs10813831) and melanoma differentiation-associated gene 5 (MDA5) (rs1990760)—with the severity of EV71 infection. Polymorphisms of candidate genes in 87 EV71-infected patients and 57 asymptomatic controls were detected. Binary logistic regression analysis revealed statistically significant differences in polymorphism of MDA5 (rs1990760) between patients with severe EV71 infection and asymptomatic controls in an additive model (OR 0.424, 95% CI 0.213-0.845, p 0.015) and a dominant model (OR 0.256, 95% CI 0.103-0.635, p 0.003). Polymorphism of MDA5 (rs1990760) (OR 0.399, 95% CI 0.199-0.798, p 0.009) was found to be associated with the severity of EV71 infection with the analysis of ordinal logistic regression. These results indicated the association between MDA5 (rs1990760) polymorphism and an increased risk of a severe EV71 infection in Chinese children, which offers potential for investigating the innate immune mechanism of EV71 infection and identifying at-risk infants, for whom a preventive strategy may reduce the severity of EV71 infection.     

Long-term follow-up of hepatitis B virus and hepatitis C virus replicative levels in chronic hepatitis patients coinfected with both viruses

Dual infection with hepatitis B and C viruses is often encountered in endemic areas of both viruses. However, understanding of the clinical and virological implications is limited. The aim of this study was to investigate the role of each virus in liver injury and the interaction between the two viruses in dual infection with hepatitis B and C viruses. Three patients who had chronic infection with both hepatitis B and C viruses were examined, and a longitudinal study of both serum hepatitis B virus DNA and hepatitis C virus RNA levels over 4 years was undertaken. The results were correlated with serum alanine aminotransferase levels. Serum alanine aminotransferase values showed a relationship with hepatitis B virus replicative levels, but not with hepatitis C virus replicative levels in all 3 patients. Serial changes of replicative levels of both viruses were studied, and it was found that hepatitis C virus replicative levels were enhanced after the decline of hepatitis B virus replication in 1 of the 3 patients. In the remaining 2 patients, a transient rise of hepatitis C virus replicative levels in association with a decrease of hepatitis B virus replication was also observed during part of the follow-up period. These findings indicate that hepatitis B virus may play a dominant etiological role in liver injury, and that a suppressive action between hepatitis B and C viruses may occur in dual infection with both viruses. © 1995 Wiley-Liss, Inc.     

Concurrent hepatitis C virus and hepatitis delta virus superinfection in patients with chronic hepatitis B virus infection.

Since hepatitis C virus (HCV) and hepatitis delta virus (HDV) are transmitted by the same routes as hepatitis B virus (HBV), simultaneous or concurrent HCV and HDV infection in patients with chronic HBV infection may occur. To test this hypothesis and to examine the clinicohistological and immunopathological presentations of such multiple hepatitis virus infections, acute and/or convalescent serum specimens from 86 patients with acute HDV superinfection were tested by enzyme immunoassay for antibodies to HCV. Of the 86 patients, 18 (20.9%) were associated with HCV infection. Although patients with early mortality cannot be evaluated by the HCV markers used in this study, the results showed that the clinical and histologic features were similar except that patients with HCV infection were older than those without HCV infection (P less than 0.01). Immunopathological studies carried out within 2 months after the onset of acute HDV superinfection demonstrated that hepatitis B core antigen (HBcAg) was not detected in any patient and HDV antigen was detected in 18.2% of the patients with HCV infection whereas HBcAg and HDAg were found in 7% and 65.1%, respectively, of those without HCV coinfection (P less than 0.02). It is concluded that concurrent HCV and HDV superinfections can and do occur in patients with chronic HBV infection. In these triple viral infections, HCV may even transiently suppress HDV and HBV.     

血清乙肝病毒大蛋白在HBeAg阴性和低水平HBV-DNA乙肝患者中的检测意义

为了探讨血清乙肝病毒大蛋白在HBeAg阴性与低水平HBV-DNA乙肝患者中的检测意义。对162例HBV感染者及47名健康对照血清采用酶联免疫吸附试验检测乙肝病毒大蛋白、乙肝病毒前S1抗原、病毒前S2抗原及乙肝病毒标志物;FQ-PCR定量检测HBV-DNA。结果显示:162例HBV感染者血清中,HBV-LP浓度与HBV-DNA拷贝数间具有良好的正相关性(rs=0.64,P<0.001),不同HBV-DNA拷贝数组别间HBV-LP浓度存在差异显著性(P<0.01);HBV-LP与HBV-DNA、HBeAg、HBVpreS2、HBVpreS1间均关联显著(P<0.01)。HBV-LP与HBV-DNA、HBeAg、HBVpreS2、HBVpreS1间阳性率均存在差异显著性(P<0.05),HBV-LP阳性率为84.57%,较HBV-DNA、HBeAg、HBVpreS2、HB-VpreS1均敏感。其中HBeAg阴性组中HBV-LP与HBVpreS1、HBVpreS2、HBV-DNA间阳性率均存在差异显著性(P<0.05),HBV-LP阳性率为76.77%(76/99),较HBV-DNA,HBVpreS2,HBVpreS1均高(P<0.01);DNA阴性组中HBV-LP与HBVpreS2、HBVpreS1、HBeAg间阳性率均存在差异显著性(P<0.05),HBV-LP阳性率为73.33%,较HB-VpreS2、HBVpreS1、HBeAg均高。血清HBV-LP浓度是反映血清HBeAg阴性和低水平DNA的HBV感染者体内病毒复制、疾病进程、疗效与预后判断的新的敏感监测指标。     

962例外来工的乙型肝炎表达模式分析

目的了解来东莞工作的外来工的乙肝感染状况，分析其乙型肝炎的表达模式，为乙肝防治提供参考。方法收集了到某院体检的962份外来工的血清，检测乙肝五项：表面抗原（HBsAg）、表面抗体（HBsAb）、e抗原（HBeAg）、e抗体（HBeAb）、核心抗体（HBcAb），并对结果进行分析。结果在检测的962人中，HBsAg阳性335人，阳性率34．82％。其中男性受检者527人，HBsAg阳性204人，阳性率38．71％；女性受检者435人，HBsAg阳性131人，阳性率30．11％。乙肝五项标志物全阴者217人，占总受检人数的22．56％。结论必须加强对来莞工作的外来工的乙肝监测，及时为乙肝五项全阴者注射乙肝疫苗，同时加强外来工的健康教育，提高他们的卫生知识水平。     

A tumour necrosis factor-alpha (TNF-α) promoter polymorphism is associated with chronic hepatitis B infection

Cytokines such as TNF-α and interferon gamma (IFN-γ) are important for the elimination of infected hepatocytes during acute hepatitis B virus (HBV) infection. Two G versus A transitions in the TNF-α promoter region at positions ?308 and ?238 possibly influence TNF-α expression. We investigated these TNF-α polymorphisms in 71 patients with chronic HBV infection, in 32 subjects that had spontaneously recovered from acute HBV infection, and in 99 healthy controls. The ?238 A promoter variant was present in 18 (25%) of 71 patients with chronic HBV infection compared with two (6%) of 32 subjects with acute infection (P < 0.04), and seven (7%) of 99 controls (P < 0.003). By contrast, the prevalence of the variant at position ?308 was similar in all investigated groups. The observed differences could not be explained by linkage disequilibrium to HLA-B or -DRB1* alleles. These findings suggest an association between the TNF-α promoter polymorphism at position ?238 and the development of chronic HBV infection. This promoter variant appears to be linked to defective viral clearance.     

Acute hepatitis B virus infection in Turkey: epidemiology and genotype distribution

The aim of this study was to investigate the prevalence of hepatitis B virus (HBV) genotypes in Turkey. Epidemiological and clinical data for 158 patients with acute HBV infection from 22 medical centres in the period February 2001 to February 2002 were collected prospectively. HBV genotyping was based on analysis of restriction fragment length polymorphisms and nested PCR. There were 59 female and 99 male patients, with a mean age of 34.2 +/- 15.6 years. The most common probable transmission route was blood contact in 63 (41.1%) cases, but was unknown in 78 (49.4%) cases. The mean alanine aminotransferase level was 1718 +/- 1089 IU/L. Four of the 158 patients (2.5%) died because of fulminant hepatitis. One year after discharge, 11 (10.6%) of 103 cases were positive for hepatitis B surface antigen (HBsAg) and 80 (77.7%) were positive for anti-HBsAg. Genotype determination was unsuccessful in 11 cases because of a negative PCR; genotype D was found in the remaining 147 cases. The results suggested that acute HBV infection constitutes a significant health problem in Turkey and that genotype D is predominant.     

Prevalence and significance of antibodies to hepatitis C virus among Saudi haemodialysis patients.

Seventy-four patients who were maintained on chronic haemodialysis in King Khalid University Hospital, Riyadh, Saudi Arabia were tested using the recently available ELISA to determine the prevalence of antibodies to hepatitis C virus (anti-HCV) in a haemodialysis unit. The prevalence rate of anti-HCV antibodies of 41.9% in the haemodialysis patients was significantly higher than the rate of 3.9% detected in 488 asymptomatic blood donors who were similarly tested. In the haemodialysis patients, anti-HCV positivity was related to previous blood transfusion (greater than 5 units of blood) and to the duration of haemodialysis (greater than 4 years); but was unrelated to sex, age, positive HBV markers or to past or current elevation of serum ALT. The results indicate a relatively higher prevalence of anti-HCV antibodies in our patients compared to rates of 1-20% reported from Europe and the U.S.A. An effective control strategy for HCV infection in this high risk group is urgently indicated.     

乙肝疫苗联合乙肝免疫球蛋白对阻断母婴垂直传播乙型肝炎病毒的临床分析

目的探讨孕妇产前用乙肝免疫球蛋白（HBIG）与乙型肝炎疫苗联合免疫阻断母婴传播的效果。方法将504例HBsAg（＋）孕妇分为A（预防组）,B（对照组）两组。A组：246名HBsAg阳性孕妇孕晚期每月分别注射基因重组型乙肝疫苗10μg、HBIG200IU（200IU/ml）,新生儿出生后采股静脉血,同时在出生后24h内注射HBIG200IU,然后在0、1、6月龄接种基因重组型乙肝疫苗,每次10μg。B组：258例产前未注射HBIG和基因重组型乙肝疫苗的HBsAg阳性孕妇,其所生新生儿在0、1、6（30μg、30μg、30μg）月龄只用基因重组型乙肝疫苗免疫。A、B两组婴儿都分别在0、3、6、9、12、24月龄静脉采血,用酶联免疫吸附试验（ELISA）检测HBV标志物,同时随访。结果A组的宫内感染率为3.25%,B组为4.16%,差异无统计学意义（χ^2=1.43,P〉0.05）。A组没有发生慢性HBV感染的婴儿,而B组中有7例婴儿发生慢性HBV感染,B组婴儿发生慢性HBV的感染率显著高于A组（χ^2=4.41,P〈0.05）。结论产前用HBIG和新生儿HBIG联合免疫可降低慢性HBV感染率,阻断宫内感染的慢性化,提高产程感染的阻断效果。     

Molecular characterization and functional analysis of occult hepatitis B virus infection in Chinese patients infected with genotype C

Occult HBV infection is defined as the persistence of HBV DNA in individuals negative for HBV surface antigen (HBsAg), and many different mechanisms have been reported in different countries. However, in China, one of the endemic areas for HBV infection, no reports have been published on occult HBV infection. The present study investigated the virological features and the mechanism of occult HBV infection in China. Full‐length HBV DNA from eight patients with occult HBV infection (S1–S8) and three HBsAg‐positive cases (SWT1–SWT3) was cloned and sequenced. Additionally, four entire linear HBV genomes from occult cases were transfected transiently into HepG2 cells. The sequencing results showed two major mutations in patients with occult HBV infection as follows: deletions in the pre‐S1 (S3, S4, and S7) and X (S1, S2, and S5) regions. Such deletions covered the S promoter and the basal core promoter (BCP), and function analysis of these variants also showed a decrease in DNA replication and antigen expression. Two patients with occult infection (S6 and S8) had no mutations capable of interfering with viral replication and gene expression in the major viral population. Thus, the deletions in the S promoter and the BCP regions that disable the regulatory elements may be the reason for the absence of HBsAg, and multiple mechanisms may be responsible for occult HBV infection. J. Med. Virol. 81:826–835, 2009. © 2009 Wiley‐Liss, Inc.     

Phylogenetic investigation of nosocomial transmission of hepatitis C virus in an oncology ward

Nosocomial hepatitis C virus (HCV) infections have been reported from different health-care settings worldwide. Twenty patients, treated at the same oncology department, with no previous record of hepatitis C infection, tested positive for anti-HCV antibodies between November 2007 and June 2008. Twelve of the newly infected patients were found to be HCV RNA positive. The common origin of the infections was assumed. To investigate the relatedness of the detected viral strains phylogenetic analyses were performed using sequences from the NS5B and E1/E2 genome regions. A patient carrying HCV for years was also involved in the study. She was treated at the same oncology department and was considered a possible infectious source. The previous HCV carrier harbored subtype 1b, while all other patients were infected with subtype 1a. Sequences from the 12 newly infected patients formed two groups. The viral sequences within the groups were very closely related. A greater evolutionary distance was observed between the two groups; however, their relatedness could be demonstrated by sequences from both regions with high statistical support. The results indicated that nosocomial transmission occurred. The phylogenetic analyses suggested that the viruses originated from a common source, possibly a patient carrying highly divergent variants. This presumed infectious source could not be identified in the course of this study. The genotype distribution of Hungarian control sequences included in the analysis confirmed this conclusion, since HCV genotype 1a was found to be relatively uncommon.     

Interference of replication between hepatitis B and C viruses in patients infected with HIV

The clinical and cellular interactions between hepatitis B virus (HBV) and hepatitis C virus (HCV) were investigated in patients co-infected with the human immunodeficiency virus (HIV). One hundred ninety-nine patients followed for 6 years were evaluated to compare the level of HBV DNA and HCV RNA in patients co-infected with HIV and HBV, and patients co-infected with HIV, HBV, and HCV. A full-length HBV genome and HCV JFH1 RNA were co-transfected into HuH-7.5.1 cells in vitro to examine the impact of co-infection and dependence on the HBV PreC mutant for replication interference. Before 2',3'-dideoxy-3'-thiacytidine (3TC)-based antiretroviral therapy (ART) was initiated, HBV DNA was found in 56/123 (45.4%) patients co-infected with HIV and HBV, and in 19/76 (25.0%) patients co-infected with HIV, HBV, and HCV. After 3TC-based ART was initiated, detectable HBV DNA decreased to 7/76 (9.2%) in patients co-infected with HIV, HBV, and HCV, but HCV RNA increased from 43/76 (56.6%) to 60/76 (78.9%) (P = 0.003). In vitro HBV and HCV co-infection led to decreased replication of both viruses. The primary factors that influenced the decreased replication were the order of the HBV and HCV infection and the HBV PreC mutation.     

急性鸭乙型肝炎病毒感染病毒清除机理研究

目的 :进一步阐明嗜肝病毒自然感染过程中病毒清除机理。方法 :7只 2～ 3月龄成年重庆麻鸭静脉接种 10～ 2 0mlDHBV阳性血清 (5× 10 8～ 1× 10 9genome) ,接种后每周采血检测外周血中DHBVDNA、DHBsAg和特异抗体的产生 ;感染后第10、35天分离外周血单个核细胞用于抗原特异细胞增殖实验 ;第 5、30、6 0天取肝组织标本进行DHBVDNASouthern杂交、DHB sAg免疫组化及肝组织病理检测。结果 :DHBV感染成年鸭在 1～ 2w潜伏期后出现急性、一过性感染 ,感染高峰期肝内存在多拷贝的DHBV所有复制中间体形式 ,包括cccDNA。进一步分析显示病毒血症的消失是在快速抗原特异细胞增殖反应和高滴度特异抗体产生之后 ;与此同时 ,整个急性感染期间 ,肝细胞并无明显的损害。结论 :非细胞直接损伤机制在嗜肝病毒清除过程中发挥了重要作用。