关于庚型肝炎病毒（GBV—C／HGV）致病性的研究进展

病毒性肝炎的病因除已知的甲、乙、丙、丁、戊型肝炎病毒，并除外其它相关病毒，如ＥＢ病毒，瘤疹病毒，尚有１０％～２０％的肝炎病因不明。１９９５年发现了一种新型肝炎病毒－庚型肝炎病毒（ＧＢＶ－Ｃ／ＨＧＶ），随着对它与肝病致病的相关性研究，ＧＢＶ－Ｃ／ＨＧＶ成为非甲、乙、丙、丁、戊型肝炎病因的一种可能的解释。ＧＢＶ－Ｃ／ＨＧＶ为单股正链 ＲＮＡ病毒基因组全长约为１０００个碱基。为了阐明ＨＧＶ的致病作用，ＨＧＶ与肝病的关系，ＨＧＶ在人体内的复制部位，以及ＨＧＶ与其它肝外疾病的关系成为研究的热点。从１９９５…     

GBV—C／HGV病毒学特征

新型病毒ＧＢＶ－Ｃ／ＨＧＶ尚未得到国际病毒分离与命名委员会的最后命名，但因其可能与人类肝炎相关，而引起国际医学界的广泛关注。本文综述了ＧＢＶ－Ｃ／ＨＧＶ病原学研究的进展，包括ＧＢＶ－Ｃ／ＨＧＶ的发现及病毒学特征研究的最新成果。     

庚型肝炎病毒C型（GBV—C）多肽的抗原性研究及应用

应用Ｇｏｌｄｋｅｙ软件分析ＧＢＶ－Ｃ的氨基酸序列，采用标准的Ｍｅｒｉｆｉｅｌｄ固相法和多中心同步多肽合成技术研究ＧＢＶ－Ｃ多肽抗原，应用抗原反应性好的ＮＳ３区Ｐ１和Ｐ１１多肽以及ＮＳ５区的Ｐ２２多肽研制抗ＧＢＶ－Ｃ检测试剂。９９例ＧＢＶ－ＣＲＮＡ阳性血清的抗ＧＢＶ－Ｃ阳性率为８１．８２％（８１／９９），１０００例单采浆献血员血样的抗ＧＢＶ－Ｃ阳性率为５．１０％（５１／１０００），抗ＧＢＶ－Ｃ阳性血样中的ＧＢＶ－ＣＲＮＡ阳性率为７０．５９％（３６／５１）。结果表明，Ｐ１，Ｐ１１和Ｐ２２可以用于抗ＧＢＶ－Ｃ检测试剂的开发。应尽快对献血员，尤其是献浆员进行抗ＧＢＶ－Ｃ的筛选。     

肝病患者中庚型肝炎病毒感染的检测

目的探讨临床肝病病人中庚型肝炎病毒（ＧＢＶ－Ｃ／ＨＧＶ）感染情况及临床特点。方法应用庚型肝炎病毒基因组５’ＵＴＲ两对寡核苷酸作为引物，建立逆转录套式聚合酶链反应，检测１６９例不同肝病患者血清标本中ＧＢＶ－Ｃ／ＨＧＶＲＮＡ，并对其中１例ＰＣＲ扩增产物进行克隆及测序。结果１６９例各型肝病病人ＧＢＶ－Ｃ／ＨＧＶＲＮＡ总的检出率为９５％（１６／１６９）。在２９例有手术输血史患者中，３１０％（９／２９）ＧＢＶ－Ｃ／ＨＧＶＲＮＡ呈阳性，明显高于无手术输血史组（５％，Ｐ＜００１）。序列分析显示１株庚肝病毒５’ＵＴＲ部分基因片段与已知庚肝病毒株核苷酸同源性在８９１４％～９８９１％之间。结论ＧＢＶ－Ｃ／ＨＧＶ感染普遍存在于临床肝病患者中，病人感染ＧＢＶ－Ｃ／ＨＧＶ的临床表现未发现有特殊性，ＧＢＶ－Ｃ／ＨＧＶ可能不是非甲～戊型肝炎的主要致病因子。     

庚型肝炎病毒的分子生物学研究进展

庚型肝炎病毒（ＧＢＶ－Ｃ／ＨＧＶ）为单股正链ＲＮＡ病毒，属黄病毒家族成员。ＧＢＶ－Ｃ／ＨＧＶ与丙型肝炎病毒（ＨＣＶ）有许多相似之处，但又有其自身特点。本文对ＧＢＶ－Ｃ／ＨＧＶ的基因组结构及变异、病毒多蛋白前体的加工、感染途径与致病性及检测方法等进行了综述，     

HCV感染外周血单个核细胞及其对T淋巴细胞亚群的影响

目的：研究丙型肝炎病毒（ＨＣＶ）感染外周血单个核细胞（ＰＢＭＣ）的情况及其对Ｔ淋巴细胞亚群的影响。方法：运用非同位素原位杂交（ＮＩＳＨ）法和链酶亲和素－生物素（ＳＡＢＣ）法分别检测２０例慢性丙型肝炎患者ＰＢＭＣ中的ＨＣＶ－ＲＮＡ和非结构（Ｎｏｎｓｔｒｕｃｔｕｒａｌ,ＮＳ）蛋白ＮＳ５抗原,同时用ＳＡＢＣ法检测其Ｔ淋巴细胞亚群。结果：８例（４０．０％）患者的ＰＢＭＣ中ＨＣＶ－ＲＮＡ呈构（Ｎｏｎｓｔｒｕ     

庚型肝炎病毒感染对慢性丙型肝炎的临床与病理学影响

目的 探讨庚型肝炎病毒（ＨＧＶ）感染对慢性丙型肝炎（ＣＨ－Ｃ）的临床与病理学影响。方法 应用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）法对５３例经肝穿活检确诊的ＣＨ－Ｃ患者血清标本进行了ＨＣＶ－ＲＮＡ检测,并将合并与未合并ＨＧＶ感染者进行了临床与病理学对比。结果 ＨＣＶ－ＲＮＡ阳性１５例（２８．３０％）。合并与未合并ＨＧＶ感染者在临床表现、肝功能等生化指标水平、ＨＣＶ－ＲＮＡ阳性率及肝脏病理损害等方面差     

表达人HGF／MBP融合蛋白的pMAL—MBP／HGF重组质粒的构建及鉴定

将人ＨＧＦ完整编码区ｃＤＮＡ片段插入ＭＢＰ表达型ｐＭＡＬ－Ｃ２载体质粒中，构建了表达人ＨＧＦ／ＭＢＰ融合蛋白的ｐＭＡＬ－ＭＢＰ／ＨＧＦ重质粒。表达的ＨＧＦ／ＭＢＰＧＥ分析分子量约为１１０ｋＤ，Ｗｅｓｔｅｒｎｂｌｏｔｔｉｎｇ表明能被兔抗ＭＢＰ抗体和抗人ＨＧＦＭｃＡｂ所识别。     

吸毒人群中HGV和HBV的同步检测

ＨＢＶ、ＨＣＶ和ＨＧＶ有共同的感染途径,ＨＢＶ为ＤＮＡ病毒,ＨＧＶ为ＲＮＡ病毒。我们建立了ＨＧＶＨＢＶＰＣＲ同步检测法,并用该方法和ＥＬＩＳＡ法检测血清中ＨＧＶＲＮＡ、ＨＢＶＤＮＡ、ＨＢｓＡｇ和抗ＨＣＶＩｇＧ以及谷丙转氨酶(ＡＬＴ)水平,调查武汉市第一戒毒所106名吸毒者ＨＧＶ与ＨＢＶ、ＨＣＶ感染情况。通过改变ＲＮＡ提取液的ｐＨ值,可同时提取ＨＧＶＲＮＡ和ＨＢＶＤＮＡ,基金项目:湖北省教委资助项目作者单位:湖北医科大学病毒研究所(郑义,伍欣星,赵,赵文先);湖北省嘉鱼县血防站(高胜君)通信作者:郑义,430022武汉市第…     

斑点杂交及RNA酶保护分析法检测反义HSP90基因转染细胞 …

目的：检测ＨＳＰ９０反义核酸转染细胞后反义ＲＮＡ的表达,方法：采用打点杂交及ＲＮＡ酶保护分析法,结果：ＨＳＰ９０反义核酸转染细胞ＡＨ－ＳＧＣ７９０１,ＡＨ－ＳＧＣ７９０１／ＶＣＲ,ＡＨ－ＨＣＣ７４０２及ＡＨ－Ｅｃ１０９有ＨＳＰ９０反义ＲＮＡ的表达。结论：ＥＳＰ９０反义ＲＮＡ在ＡＨ－ＳＧＣ７９０１,ＡＨ－ＳＧＣ７９０１／ＶＣＲ,ＡＨ－ＨＣＣ７４０２及ＡＨ－Ｅｃ１０９细胞的表达,为进一步研究ＨＳＯ９０     

GBV-C/HGV infection in chronic hepatitis C patients: Its effect on clinical features and interferon therapy

A novel virus (GBV-C/HGV) may be associated with some liver diseases including fulminant hepatitis and acute and chronic hepatitis. On the other hand, many investigations showed that this infection does not contribute to liver disease. GBV-C/HGV has been found to occur in association with infection with other hepatitis viruses. We investigated the effect of GBV-C/HGV infection on the clinical features and interferon treatment in patients with chronic hepatitis C. A total of 262 hepatitis C virus (HCV) RNA positive patients with chronic hepatitis were examined in this study. The detection of serum GBV-C/HGV RNA was done by RT-PCR using specific primers from the NS5 regions. Interferon-alpha was given at a dose of 6 MU/day for 16 or 24 weeks. A responder was defined as a patient with ALT normalization and HCV RNA disappearance after treatment. GBV-C/HGV RNA was detected in 28 (11%) patients. No significant difference was detected in clinical features (age, sex, liver-related biochemical tests, and histological examination) between the 28 GBV-C/HGV-positive patients and the GBV-C/HGV-negative patients. Using interferon therapy for hepatitis C, the responder rates of GBV-C/HGV-positive and -negative patients were 14% and 20%, respectively. Of the 28 patients with GBV-C/HGV RNA, GBV-C/HGV RNA was tested after interferon therapy in 16 and of these GBV-C/HGV RNA was not detected in nine patients after therapy. These findings suggest that GBV-C/HGV infection dose not affect the clinical features in patients with HCV and the efficacy of interferon therapy for chronic hepatitis C. J. Med. Virol. 55:98–102, 1998. © 1998 Wiley-Liss, Inc.     

Identification of GBV-C hepatitis G RNA in chronic hepatitis C patients

Sera from patients with chronic hepatitis C were examined for the presence of GBV-C/HGV RNA by RT-PCR. The amplified products, derived from the 5′ non-coding, NS3, and NS5a regions, were detected in 19 (19%) of the 100 HCV RNA-positive samples. Analysis of GBV-C/HGV prevalence rates revealed that dual infections are related to shared parenteral risk factors. Intravenous drug abuse and multiple transfusions were the factors clearly associated with a simultaneous HCV and GBV-C/HGV infection. Apart from this, patients with dual infections had a statistically significant lower mean age compared to those patients infected solely by HCV. Determination of HCV genotypes involved in GBV-C/HGV coinfection by RFLP analysis showed no correlation between the presence of GBV-C/HGV and a distinct HCV genotype. The study demonstrates that, based on the assessment of risk criteria, GBV-C/HGV is transmitted efficiently parenterally and is frequently linked to hepatitis C coinfection, regardless of HCV genotype. © 1996 Wiley-Liss, Inc.     

Chronic hepatitis C infection: influence of the viral load, genotypes, and GBV-C/HGV coinfection on the severity of the disease in a Brazilian population

The distributions of the different genotypes of the hepatitis C virus (HCV) and GBV-C virus (GBV-C/HGV) vary geographically and information worldwide is still incomplete. In particular, there are few data on the distribution of genotypes (and their relationship to the severity of liver disease) in South America. Findings are described in 114 consecutive patients from Northeast Brazil (median age 52 years, range 18-72 years) who had abnormal levels of serum aminotransferases and seropositivity for HCV RNA. The patients were recruited from an outpatient clinic between November 1997 and April 1998. Quantitative HCV RNA and GBV-C/HGV RNA estimations were carried out by double-nested polymerase chain reaction (PCR) using primers from the 5'-untranslated regions (UTRs) of the genomes. HCV genotypes were determined by restriction fragment length polymorphism (RFLP) analysis with 5'-UTR primers and by PCR with type-specific 5'-UTR primers. GBV-C/HGV-RNA genotypes were determined by RFLP with specific 5'-UTR primers and phylogenetic trees were constructed using the Neighbour-Joining and Drawtree programs. Histological features were graded and staged according to international criteria. Of the 114 patients, 35 (30.7%) patients had cirrhosis and 22 (27.8%) had mild, 51 (64.6%) had moderate, and 6 (7.6%) had severe chronic hepatitis. Median HCV viral load was 10(6) genome equivalents per millilitre (range 10(4)-10(9)/ml). Frequencies of genotypes were 5.3% type 1a, 44.7% type 1b, 3.5% type 2, 41.2% type 3, and 5.3% mixed types. GBV-C/HGV-RNA was detected in the sera of 12 (10.5%) patients and was distributed among three phylogenetic groups. There were no significant differences between patients with the predominant HCV genotypes (1b and 3) with respect to gender, age group, viral load, severity of liver disease, or coinfection with GBV-C/HGV.     

Correlation of interferon treatment response with GBV-C/HGV genomic RNA and anti-envelope 2 protein antibody

The clinical significance of GB virus C/hepatitis G virus (GBV-C/HGV) co-infection was studied retrospectively in 100 consecutive patients with hepatitis C virus (HCV) infection. All 100 patients had been treated with interferon-alpha (IFN-alpha). Co-infection with GBV-C/HGV and HCV was detected in 10 of the 100 patients (10%) and anti-envelope 2 region (anti-E2) antibody was detected in 25 patients. None of the patients with GBV-C/HGV RNA had anti-E2 antibody. Co-infected patients were younger (P < .005) and their serum transaminase levels were lower than HCV-only infected patients (P< .01). In 7 of the 10 co-infected patients, HCV RNA was eradicated from serum after IFN-alpha treatment and normal alanine transaminase (ALT) levels continued in 6 of these 7 patients. In one patient who was negative for HCV RNA but positive for GBV-C/HGV RNA, the ALT level relapsed transiently. The rate of clearance of HCV and normalization of the ALT level was significantly higher in co-infected patients than in HCV-only infected patients (P < .05). GBV-C/HGV RNA disappeared from 6 of the 10 co-infected patients (60%) upon cessation of IFN-alpha treatment. However, continuous clearance of GBV-C/HGV was observed in only two patients and anti-E2 antibody could not be detected in the serum of these patients. These results indicate that co-infected patients tend to be younger and more sensitive to IFN-alpha treatment. However, long-term clearance of GBV-C/HGV after IFN-alpha treatment may be difficult. Moreover, anti-E2 antibody may act to neutralize GBV-C/ HGV.     

Prospective follow-up of patients with GBV-C/HGV infection: specific mutational patterns, clinical outcome, and genetic diversity

An association between a specific mutational pattern within the nonstructural (NS)3 region of GB virus-C/hepatitis G virus (GBV-C/HGV) genome and fulminant hepatic failure has been suggested recently. The mutational pattern consists of 3-6 nucleotide mutations of which one is leading to an amino acid exchange. In the present study, patients with GBV-C/HGV mono-infection (n = 24) or GBV-C/HGV and HCV co-infection (n = 20) were investigated prospectively. In 6/44 patients (14%) the mutational pattern within GBV-C/HGV NS3 previously associated with fulminant hepatic failure was identified by direct sequence analysis of the NS3 region. All 44 patients were asymptomatic clinically and had normal liver functions at initial presentation and after a median follow-up of 2.2 years. In 22/24 patients with GBV-C/HGV mono-infection and all patients with GBV-C/HGV and HCV co-infection GBV-C/HGV RNA remained detectable at the end of the study period, whereas two patients infected with GBV-C/HGV alone became negative for GBV-C/HGV RNA and developed GBV-C/HGV anti-E2 antibodies indicating recovery from GBV-C/HGV infection. Aminotransferase levels remained elevated or became normal independent of the persistence of serum GBV-C/HGV RNA. The median rate of nucleotide substitutions in GBV-C/HGV mono-infected and HCV co-infected patients was 3.4 x 10(-3) and 3.2 x 10(-3) per site per year, respectively. In conclusion, the prevalence of the mutational pattern within NS3 region of GBV-C/HGV associated previously with fulminant hepatic failure is about 14% and not associated specifically with severe liver disease. Over a median follow-up of 2.2 years less than 5% of patients cleared spontaneously GBV-C/HGV and no correlation between viraemia and elevated liver enzymes was observed.     

Direct evidence for GB virus C/hepatitis G virus (GBV-C/HGV) superinfection: elimination of resident viral strain by donor strain in a patient undergoing liver transplantation

The viral genome of GB virus C/hepatitis G virus (GBV-C/HGV), a single-strand RNA virus, is subject to considerable variability and at least four genotypes have been suggested based on phylogenetic analysis. While co-infection of GBV-C/HGV with other infectious agents such as hepatitis C virus (HCV) has been frequently observed, there is no report whether or not co-infection and/or superinfection occurs among different GBV-C/HGV strains. By studying a GBV-C/HGV positive recipient/donor pair in the context of undergoing liver transplantation, we have sequenced multiple clones derived from serum samples serially collected over four years. Detailed phylogenetic analyses have been performed with these sequences. The donor was infected with GBV-C/HGV genotype 1 and this strain completely replaced recipient GBV-C/HGV strain (genotype 2) after liver transplantation. The recipient's original viral strain became undetectable during follow-up. Sequence analysis failed to identify genetic recombination between the two genotypes, at least in whole structural domain. This study, therefore, provides direct evidence for GBV-C/HGV superinfection of one strain by another with one of them predominating probably due to replication competition.     

GBV-C/HGV infection in hepatitis C virus-infected deferred Swedish blood donors

Sera from 62 hepatitis C virus (HCV)-infected Swedish blood donors were tested by a nested polymerase chain reaction using primers targeting the 5′-noncoding region of the GB virus-C/hepatitis G (GBV-C/HGV) genome and an enzyme-linked immunosorbent assay that detects antibodies to the envelope protein E2 of GBV-C/HGV (anti-E2). Fourteen (22%) and 21 (34%) of the 62 blood donors were found to be GBV-C/HGV RNA and anti-E2 positive, respectively. None of the blood donors was positive for both GBV-C/HGV RNA and anti-E2. Thus, 35 of 62 (56%) HCV-infected donors had been exposed to GBV-C/HGV infection. At sequencing of the 14 GBV-C/HGV isolates, 12 were identified as subtype 2a and 2 as subtype 2b. One of 7 (14%) donors with mild liver disease such as steatosis and nonspecific reactive hepatitis had been exposed to GBV-C/HGV vs. 34 of 55 (62%) with chronic hepatitis with or without cirrhosis (P = 0.04). All other differences in histology were small between HCV and dual HCV GBV-C/HGV-infected donors. In conclusion, more than half of HCV-infected Swedish blood donors in this study were positive for either GBV-C/HGV RNA or anti-E2. GBV-C/HGV viremia and seropositivity were mutually exclusive. J. Med. Virol. 54:75–79, 1998. © 1998 Wiley-Liss, Inc.