HCV不同功能区抗体与HCVRNA的关系探讨

探讨HCV不同功能区抗体(抗-C、抗-NS3、抗-NS4及抗-NS5)与HCVRNA的关系。HCV不同功能区抗体测定采用ELISA法,用RT-PCR进行HCV RNA检测。结果显示55例抗HCV阳性血清中有7种抗体阳性组合。抗-NS3、抗-C、抗-NS5及抗-NS4在7种阳性组合中的检出率分别为96．4%、89．1%、58．2%、56．4%。HCVRNA阳性血清中抗-NS3、抗-C、抗-NS5及抗NS4的检出率分别为96．2%,88．5%,57．7%,57．7%。HCV不同功能区抗体ELISA法有很高的敏感性和特异性,与RT-PCR法基本一致。抗-NS3、抗-C在HCV的诊断中有重要的意义,抗-NS5和抗-NS4有诊断的互补作用。     

不同HCV感染者各区段抗HCV抗体的反应性及HCV-RNA检测结果分析

利用丙型肝炎病毒（ＨＣＶ）不同区段抗原的单片段酶免疫（ＥＩＡ）试剂及反转录聚合酶链反应（ＲＴ－ＰＣＲ）技术,对不同ＨＣＶ感染者血清不同区段抗ＨＣＶ抗体及丙型肝炎病毒核糖核酸（ＨＣＶ－ＲＮＡ）进行检测。结果显示,慢性肝炎患者各抗体检出率及Ｓ／Ｃｏ（Ｓ为样品光密度值,Ｃｏ为临界值）值均高,肝癌患者各抗体检出率及ＨＣＶ－ＲＮＡ阳性率均较低;ＡＬＴ正常而抗ＨＣＶ阳性者各区段抗体阳性率低于急、慢性肝炎患者,而其ＨＣＶ－ＲＮＡ的阳性率高于其他各组;抗－ＮＳ３和抗－Ｃ抗体在各组的检出率和反应性均强。表明抗－Ｃ和抗－ＮＳ３抗体的检测对ＨＣＶ感染最有诊断价值,同时用ＲＴ－ＰＣＲ技术检测血清ＨＣＶ－ＲＮＡ有助于ＨＣＶ感染的早期诊断     

HCV reinfection incidence among individuals treated for recent infection

One challenge to HCV elimination through therapeutic intervention is reinfection. The aim of this analysis was to calculate the incidence of HCV reinfection among both HIV‐positive and HIV‐negative individuals treated for recent HCV infection (estimated infection duration <18 months). Individuals with recent HCV infection who achieved an end‐of‐treatment response in four open‐label studies between 2004 and 2015 in Australia and New Zealand were assessed for HCV reinfection, confirmed by sequencing of the Core‐E2 and/or NS5B regions. Reinfection incidence was calculated using person‐time of observation. Exact Poisson regression analysis was used to assess factors associated with HCV reinfection. The cohort at risk for reinfection (n=120; 83% male; median age 36 years) was composed of HIV‐positive men‐who‐have‐sex‐with‐men (53%) and people who inject drugs (current 49%, ever 69%). Total follow‐up time at risk was 135 person‐years (median 1.08 years, range 0.17, 2.53). Ten cases of HCV reinfection were identified, for an incidence of 7.4 per 100 py (95% CI 4.0, 13.8). Reinfection incidence was significantly higher among participants who reported injection drug use at end of or post‐treatment, irrespective of HIV status (15.5 per 100 py, 95% CI 7.8, 31.1). In adjusted analysis, factors associated with reinfection were older age (aIRR 5.3, 95% CI 1.15, 51.5, P=.042) and injection drug use at end of or post‐treatment (aIRR 7.9, 95% CI 1.6, 77.2, P=.008). High reinfection incidence following treatment for recent HCV infection in individuals with ongoing risk behaviour emphasizes the need for post‐treatment surveillance, harm reduction strategies and education in at‐risk populations.     

HCV genotypes in hepatitis C patients and their clinical significances

ＩＮＴＲＯＤＵＣＴＩＯＮＩｔｈａｓｂｅｅｎｄｉｓｃｏｖｅｒｅｄｔｈａｔｈｅｐａｔｉｔｉｓＣｖｉｒｕｓ（ＨＣＶ）ｐｒｅｓｅｎｔｓｃｏｎｓｉｄｅｒａｂｌｅｎｕｃｌｅｏｔｉｄｅｖａｒｉａｔｉｏｎａｎｄｈａｓｍａｎｙｇｅｎｏｔｙｐｅｓ．Ａｔｌｅａｓｔｔｗｅｌｖｅ,５ｏｆｗｈｉｃｈｐｒｅｖａｌｅｎｔ,ｉ．ｅ．：Ⅰ／１ａ,Ⅱ／１ｂ,Ⅲ／２ａ,Ⅳ／２ｂ,ａｎｄⅤ／３ａｔｙｐｅｓ．ＴｈｅｄｉｆｆｅｒｅｎｔｇｅｎｏｔｙｐｅｓｏｆＨＣＶｍａｙｐｏｓｓｅｓｓｓｏｍｅｒｅｔａｔｉｏｎｓｈｉｐｗｉｔｈｒｅｇｉｏｎａｌｄｉ…     

Anti‐envelope antibody responses in highly exposed seronegative individuals may be associated with protection from HCV infection

In rare cases, individuals with a history of long‐term injecting drug use remain seronegative and aviraemic, despite prolonged and likely repeated exposure to Hepatitis C virus (HCV) through high‐risk behaviour. We describe anti‐HCV Envelope (E) antibody responses in a prospective cohort of carefully defined highly exposed but uninfected subjects (HESN) and comparison subjects who were also high risk and uninfected, but rapidly became HCV infected (Incident). Longitudinally collected samples from HESN cases (n = 22) were compared to Incident controls (n = 22). IgG, IgM and IgA from sera were tested by ELISA to genotype 1a and 3a E glycoproteins, and recombinant genotype 1a E2 antigen. IgG subclass isotyping was performed for those positive for IgG. Virus‐neutralizing activity was assessed on HCV pseudoparticles, and HCV E–specific B cells analysed using flow cytometry. A significant minority of HESN cases (n = 10; 45%) had anti‐E, predominantly in the IgG2 subclass, which was not found in the pre‐infection time point of the Incident cases (n = 1; 5%). A subset of the HESN subjects also had neutralizing activity and HCV‐specific B cells detected significantly more than Incident cases pre‐infection. In conclusion, the HESN phenotype is associated with IgG2 anti‐E antibodies, neutralization activity and HCV E–specific memory B cells. These findings suggest that HESN subjects may be resistant to HCV infection through humoral immune‐mediated mechanisms.     

The epidemiologic feature of HCV prevalence in Fujian

AIM To investigate the epidemiological features of HCV prevalence, a seroepidemiological survey on HCVinfection has been carried out in Fujian since 1992.METHODS Using stratified multistage random cluster sampling, 3809 serum samples collected from 1237families in the diseases surveillance points were tested by UBI HCV EIA kit.RESULTS The results showed that the prevalence rate was 3.99%. The rate in male and female was3.63% and 4.25%, and in urban and rural 3.12% and 4.6% respectively (P>0.05). There was lower ratein children aged under 10 years. The highest rate was in 20 - 24 years old. The rates in different areas wereranged from 1.39% to 6.08% (P<0.05). The intrafamilial transmission was not important, indicating nointrafamilial aggregation. The superinfection of HCV with HAV, HBV and HEV were existed. The HCVinfection was slightly correlated with the history of hepatitis and transfusion.CONCLUSION It suggests that the HCV transmission among the population in Fujian is mainly sporadicinfection.     

HBV/HCV重叠感染的研究进展

HBV是一种嗜肝DNA病毒,HBV DNA和HBV特异P蛋白由核壳包裹成为核心颗粒,再由脂蛋白外膜包裹成完整的病毒颗粒。HCV是黄病毒科病毒,为单股正链RNA。HBV和HCV均由肠道外途径传播,2种病毒常可由相同途径发生感染。归纳了HBV/HCV重叠感染的发病机制、与隐匿性HBV感染和肝细胞癌以及器官移植、HBV疫苗之间的关系,同时介绍了重叠感染的治疗。指出存在于患者体内的HBV和HCV在病毒学方面相互干扰,在病变方面相互叠加。     

Enhanced antiviral effect in cell culture of type 1 interferon and ribozymes targeting HCV RNA

We have recently shown that the replication of an HCV-poliovirus (PV) chimera that is dependent upon the hepatitis C virus (HCV) 5' untranslated region (UTR) can be inhibited by treatment with ribozymes targeting HCV RNA. To determine the antiviral effects of anti-HCV ribozyme treatment in combination with type 1 interferon (IFN), we analysed the replication of this HCV-PV chimera in HeLa cells treated with anti-HCV ribozyme and/or IFN-α2a, IFN-α2b, or consensus IFN. The anti-HCV ribozyme, or any of the IFNs alone have significant inhibitory effects on HCV-PV replication compared to control treatment (≥ 85%, P  < 0.01). The maximal inhibition due to IFN treatment (94%, P  < 0.01) was achieved with ≥ 50 U/ml for either IFN-α2a or IFN-α2b compared to control treatment. A similar level of inhibition in viral replication could be achieved with a 5-fold lower dose of IFN if ribozyme targeting the HCV 5' UTR was given in combination. For consensus IFN, the dose could be reduced by > 12.5-fold if ribozyme targeting the HCV 5' UTR was given in combination. Conversely, the dose of ribozyme could be reduced 3-fold if given in combination with any of the IFN preparations. Moreover, treatment with low doses (1–25 U/mL) of IFN-α2a, IFN-α2b, or consensus IFN in combination with anti-HCV ribozyme resulted in > 98% inhibition of HCV-PV replication compared to control treatment ( P  < 0.01). These results demonstrate that IFN and ribozyme each have a beneficial antiviral effect that is augmented when given in combination.     

HCV对HIV/HCV共感染患者病情进展的影响

目的探讨HCV对HIV/HCV共感染病情进展的影响。方法研究对象为2012年8月到北京佑安医院随访的HIV/HCV共感染者29例及HIV单独感染者20例。两组患者年龄、性别及HIV感染时间及感染方式、感染的HIV病毒亚型均具有可比性。外周血生化指标检测并采用瞬时弹性扫描仪FibroScan评估肝脏功能及纤维化程度,运用流式细胞技术检测外周血CD4+T、CD8+T细胞绝对计数。两组计量资料比较采用t检验,计数资料比较采用χ2检验。结果 HIV/HCV共感染组ALT、AST及TBil水平分别为(76.16±81.25)U/L、(87.66±71.32)U/L、(14.21±9.56)μmol/L,明显高于HIV单独感染组[(27.74±20.63)U/L、(45.65±16.95)U/L、(10.26±3.22)μmol/L],差异具有统计学意义(P值分别为0.004、0.005及0.046)。与HIV单独感染组相比,HIV/HCV共感染组Stiffness指数有升高的趋势,但差异无统计学意义(t=1.889,P=0.080)。HIV/HCV共感染组HIV病毒载量(拷贝/ml)的对数值为3.66±0.97明显高于HIV单独感染组的3.02±0.90(t=2.251,P=0.030)。HIV/HCV共感染组、HIV单独感染组CD4+T淋巴细胞计数及CD4+T/CD8+T细胞比例分别为(374.25±185.48)/μl及(0.33±0.17)、(496.45±230.98)/μl及(0.46±0.27),HIV/HCV共感染组CD4+T淋巴细胞计数及CD4+T/CD8+T细胞比例低于HIV单独感染组,差异具有统计学意义,P值分别为0.048、0.043。共感染组艾滋病发病率(27.59%)呈现出较HIV单独感染组(5%)高的趋势(P=0.063)。结论HCV促进HIV/HCV共感染者肝脏损伤,增强HIV复制,加剧机体免疫功能损伤,HCV可能加速HIV/HCV共感染者的病情进展。     

慢性丙型肝炎病毒感染者外周血淋巴细胞增殖反应

目的　观察慢性丙型肝炎患者外周血淋巴细胞对丙型肝炎病毒（ＨＣＶ） 抗原刺激的增殖反应．方法　外周血单个核细胞（ＰＢＭＣ） 与ＨＣＶ 抗原ｃ２２ 、ｃ３３ 、ｃ１００ － ３ 、ＮＳ５ 和植物血凝素（ＰＨＡ） 分别共同孵育,加入胸腺嘧啶核苷（３ ＨＴｄＲ） ,然后收集细胞于液闪仪测定每分钟脉冲数（ｃｐｍ） ．结果　根据对不同ＨＣＶ 抗原的淋巴细胞增殖反应发现,以ｃ２２免疫原性最强,ｃ１００ － ３ 次之：淋巴细胞激活与ＨＣＶ 基因型关系不大;健康对照和慢性乙型肝炎患者对各ＨＣＶ 抗原未能显示有效的淋巴细胞增殖反应;与健康对照比较,慢性丙型肝炎和乙型肝炎患者对ＰＨＡ 刺激的淋巴细胞增殖反应降低．结论　ＨＣＶ 抗原ｃ２２ 免疫原性最强,丙型肝炎患者对ＨＣＶ 抗原的淋巴细胞增殖反应系特异性;慢性丙型肝炎和乙型肝炎患者存在抑制的细胞免疫应答．     

Effects of HCV proteins in current HCV transgenic models

Hepatits C virus (HCV) is an enveloped virus with positive‐sense single‐stranded RNA genome that causes both acute and persistent infections associated with chronic hepatitis, cirrhosis and hepatocellular carcinoma, which needs fully functional human hepatocytes for its development. Due to the strict human tropism of HCV, only human and higher primates such as chimpanzees have been receptive to HCV infection and development, cognition about pathophysiololgy and host immune responses of HCV infection is limited by lacking of simple laboratory models of infection for a long time. During the past decade, gene transfer approaches have been helpful to the understanding of the molecular basis of human disease. Transgenic cell lines, chimeric and transgenic animal models were developed and had been demonstrated their invaluable benefits. This review focuses on the existing HCV transgenic models and summarize the relative results about probable pathophysical changes induced by HCV proteins.     

Real‐world utility of HCV core antigen as an alternative to HCV RNA testing: Implications for viral load and genotype

Following positive serology, the gold standard confirmatory test of hepatitis C virus (HCV) infection is detection of HCV RNA by PCR. We assessed the utility of HCV core antigen testing to identify active infection among those positive for anti‐HCV antibodies, when introduced to routine testing. We identified serum samples that were tested at a single laboratory in Scotland from June 2011to December 2017. Serum samples testing positive for HCV antibodies (HCV Ab positive) followed by reflex HCV core antigen (Ag) testing during the study period were identified. Those patients for whom a PCR test was requested on the baseline sample were also identified. For this group, the sensitivity and specificity of HCV Ag as a diagnostic tool were assessed using HCV PCR as gold standard. In our cohort of 744 patients, we demonstrated a sensitivity of 82.1% (95% CI 77.1%‐86.2%) and a specificity of 99.8% (95% CI 98.6%‐100%). Genotype 3 was associated with increased odds of a false‐negative result (OR = 3.59, 95% CI: 1.32‐9.71), and reduced odds of a false negative were associated with older age (odds ratio (OR)=0.92, 95% CI: 0.88‐0.97 per year) and viral load (OR = 0.10, 95% CI: 0.05‐0.21 per log₁₀ IU/ml). While the implementation of HCV core antigen testing for diagnosis could lead to significant cost savings in national screening programmes, our data suggest that a significant proportion of HCV‐infected individuals may be missed. These findings have implications for HCV diagnosis and determination of viral clearance after treatment, particularly in low‐ and middle‐income regions, where genotype 3 is prevalent.     

HCV相关HCC中伴随HBV感染的意义

目的了解HCV相关HCC中伴随HBV感染的意义,初步探讨HCV和HBV在HCC的发生中的独立作用或相互作用的机制.方法原位杂交检测HCC中c-myc,H-ras,N-ras和nm23H1的mRNA表达,观察它们在伴随HBV感染组(n=44)与单纯HCV感染组(n=10)之间的差异.结果 H-ras和nm23H1在两组的表达强度有差别,H-ras在C+B组的表达强于C组(P=0.0254),而nm23H1的表达C组强于C+B组(P=0.0158).结论 HBV与HCV能协同影响H-ras的表达;单纯HCV相关HCC组之nm23H1 mRNA表达高于伴随HBV感染组,可能是HCV相关HCC低转移率的机制之一.