Distribution of different hepatitis C virus genotypes in patients with hepatitis C virus infection

AIM:To investigate the presence of mixed infection and discrepancy between hepatitis C virus(HCV) genotypes in plasma,peripheral blood mononuclear cells(PBMCs),and liver biopsy specimens.METHODS:From September 2008 up to April 2009,133 patients with chronic hepatitis C referred to Firouzgar Hospital for initiation of an antiviral therapy were recruited in the study.Five milliliters of peripheral blood was collected from each patient and liver biopsy was performed in those who gave consent or had indications.HCV genotyping was done using INNO-LiPATM HCV in serum,PBMCs,and liver biopsy specimens and then conf irmed by sequencing of 5'-UTR fragments.RESULTS:The mean age of patients was 30.3 ± 17.1 years.Multiple transfusion was seen in 124(93.2%) of patients.Multiple HCV genotypes were found in 3(2.3%) of 133 plasma samples,9(6.8%) of 133 PBMC samples,and 8(18.2%) of 44 liver biopsy specimens.It is notable that the different genotypes found in PBMCs were not the same as those found in plasma and liver biopsy specimens.CONCLUSION:Our study shows that a signif icant proportion of patients with chronic hepatitis C are affected by multiple HCV genotypes which may not be detectable only in serum of patients.

Is chronic hepatitis C virus infection a risk factor for breast cancer?

AIM:To evaluate the prevalence of breast tumors in adult females with chronic hepatitis C virus(HCV) infection.METHODS:Prospective,single-center study,based on female outpatients consulting in a liver unit,for 1 year.The study group included females with present and/or past history of chronic infection by HCV.Patients with spontaneous recovery were excluded.Chronic hepatitis had been proved by liver biopsy in the majority of cases and/or biological markers of inflammation and fibrosis.The control group included female patients with other well documented chronic liver diseases:chronic hepatitis B,alcoholic liver disease,autoimmune hepatitis,hemochromatosis,non alcoholic liver disease,chronic cholangitis.Participating patients were prospectively questioned during consultation about past breast history and follow-up by mammography.RESULTS:Breast carcinoma was recorded in 17/294 patients with HCV infection(5.8%,95% CI:3.1-8.4) vs 5/107 control patients(4.7%,95% CI:0.67-8.67).Benign tumors of the breast(mastosis,nodules,cysts) were recorded in 75/294 patients with HCV infection(25.5%,95% CI:20.5-30.5) vs 21/107(19.6%,95% CI:12.1-27.1) in the control group.No lesion was noted in 202 patients with HCV(68.7%,95% CI:63.4-74) vs 81 control patients(75.7%,95% CI:67.6-83.8).Despite a trend to an increased prevalence in the group with HCV infection,the difference was not significant compared to the control group(P=NS).In patients over 40 years,the results were,respectively,as follows:breast cancer associated with HCV:17/266 patients(6.3%,95% CI:3.4-9.3) vs 5/95 patients(5.2%,95% CI:0.7-9.7) in the control group;benign breast tumors:72/266 patients with HCV infection(27%,95% CI:21.7-32.4) vs 18/95 patients(18.9%,95% CI:11-26.8) in the control group;no breast lesion 177/266(66.5%,95% CI:60.9-72.2) in patients with HCV infection vs 72/95(75.7%,95% CI:67.1-84.4) in the control group.The differences were not significant(P=NS).CONCLUSION:These results suggest that chronic HCV infection is not a strong promoter of breast carcinoma in adult females of any age.

Recently acquired hepatitis C virus infection among people living with human immunodeficiency virus at a university hospital in Taiwan

BACKGROUND Little is known about the engagement in hepatitis C virus(HCV)care and completion of HCV treatment in people living with human immunodeficiency virus(HIV)(PLWH)who have HCV coinfection in the Asia-Pacific region.Examining the HCV care cascade can identify barriers to the completion of HCV treatment and facilitate achievement of HCV micro-elimination in PLWH.AIM To investigate the care cascade of incident HCV infections among PLWH in Taiwan.METHODS PLWH with incident HCV infections,defined as HCV seroconversion,were retrospectively identified by sequential anti-HCV testing of all archived blood samples at National Taiwan University Hospital between 2011 and 2018.All PLWH with incident HCV infections were followed until December 31,2019.The care cascade of HCV examined included all incident HCV-infected patients,the percentages of anti-HCV antibodies detected by HIV-treating physicians in clinical care,plasma HCV RNA load tested,HCV RNA positivity diagnosed,referral to treatment assessment made,anti-HCV treatment initiated,and sustained virologic response achieved.Those who had HCV seroconversion during the interferon(IFN)era(2011–2016)and the direct-acting antiviral(DAA)era(2017–2018)were analyzed separately.The duration of HCV viremia—from the date of seroconversion to viral clearance by treatments or until the end of observation—and the incidence of sexually transmitted infections(STIs)during the HCV viremic period were estimated.RESULTS During the study period,287 of 3495(8.2%)PLWH(92.3%being men who have sex with men)who were HCV-seronegative at baseline developed HCV seroconversion by retrospective testing of all archived blood samples.Of the 287 incident HCV infections,277(96.5%)had anti-HCV antibodies detected by HIV-treating physicians,270(94.1%)had plasma HCV RNA determined and 251(87.5%)tested positive for HCV RNA.Of those with HCV viremia,226(78.7%)were referred to treatment assessment,215(74.9%)initiated anti-HCV treatment,and 202(70.4%)achieved viral clearance.Compared with that in the IFN era,the median interval from HCV seroconversion by retrospective testing to detection of HCV seropositivity by HIV-treating physicians was significantly shorter in the DAA era{179 d[interquartile range(IQR)87-434]vs 92 d(IQR 57-173);P<0.001}.The incidence rate of STIs in the DAA vs the IFN era was 50.5 per 100 person-years of follow-up(PYFU)and 38.5 per 100 PYFU,respectively,with an incidence rate ratio of 1.31(95%confidence interval 0.96-1.77),while the duration of HCV viremia was 380 d(IQR 274-554)and 735 d(IQR 391-1447)(P<0.001),respectively.CONCLUSION While anti-HCV therapies are effective in achieving viral clearance,our study suggests more efforts are needed to expedite the linkage of PLWH diagnosed with incident HCV infections to HCV treatment.

Comparative Assay of Hepatitis B and C Virus Infection Markers by Different Assay Kits^1

In order to compare sensitivity of EIA and RIA assay kits for hepatitis B and C virus (HBV and HCV, respectively) infection markers, 100 serum samples in total were collected form 50 adult women each in urban and rural areas in northeast China. The number of positive cases to the three infection markers on HBV (i.e., HBsAg +, anti HBs +, and anti HBc +) and the one on HCV (anti HCV +) were examined in two laboratories, i.e., in Laboratory A with EIA kits produced in China and in Laboratory B with RIA kits. HCV infection positivity (anti HCV +) was examined by EIA kits in both laboratories, but from different sources in and outside of China, respectively. The assay in Laboratory A gave 2 HBsAg + cases out of the 100 cases examined, whereas there were 9 positive cases in Laboratory B. In contrast, 19 cases were positive to anti HCV when examined in Laboratory A, and there were 3 cases in Laboratory B. Thus, the kits used in Laboratory A gave fewer HBsAg + and more anti HCV + cases than the kits used in Laboratory B. The prevalence of anti HBs + or anti HBc + and cases did not differ when assayed in the two laboratories with EIA and RIA kits, respectively. The agreement of positive and negative findings between the two sets of testing were 93%, 93%, 93%, 86% and 82% for HBsAg, anti HBs, anti HBc, HBV (i.e., either positive to anyone of the three markers or negative to all three markers), and anti HCV, respectively. The implication of the observation on epidemiology on HBV and HCV infection prevalence was discussed.

Hepatitis C virus micro-elimination:Where do we stand?

Hepatitis C virus (HCV) elimination by 2030, using direct-acting antiviraltreatments, has been promoted by the World Health Organization. Thisachievement is not attainable, however, particularly after the 2020 pandemic ofthe coronavirus disease 2019. Consequently, the more realistic objective ofeliminating HCV from population segments for which targeted strategies ofprevention and treatment are easily attained has been promoted in Europe, as avalid alternative. The underlying idea is that micro-elimination will ultimatelylead to macro-elimination. The micro-elimination strategy may target differentspecific populations and at-risk groups. Different settings, including prisons andhospitals, have also been identified as micro-elimination scenarios. In addition,dedicated micro-elimination strategies have been designed that are tailored at thegeographical level according to HCV epidemiology and individual country’sincome. The main elements of a valid and successful micro-elimination project arereliable epidemiological data and active involvement of all the stakeholders.Community involvement represents another essential component for a successfulprogram.

Hepatocellular carcinoma after direct-acting antiviral hepatitis C virus therapy:A debate near the end

Direct acting antivirals(DAAs)have revolutionized the treatment of hepatitis C virus(HCV)infection,achieving high rates(≥95%)of sustained virological response,with a good safety profile and high compliance rates.Consequently,it had been expected that viral clearance will reduce morbidity and mortality rates,as well as the risk of hepatocellular carcinoma(HCC).However,since 2016,concerns have been raised over an unexpected high rate of HCC occurrence and recurrence after DAA therapy,which led to an avalanche of studies with contradictory results.We aimed to review the most recent and relevant articles regarding the risk of HCC after DAA treatment and identify the associated risk factors.

Monitoring hepatitis C virus treatment rates in an Opioid Treatment Program:A longitudinal study

BACKGROUND Direct-acting antivirals(DAAs)are recommended for the treatment of hepatitis C virus(HCV)infection in patients treated with methadone or buprenorphine.AIM To assess HCV treatment rates in an Opioid Treatment Program(OTP).METHODS This longitudinal study included 501 patients(81.4%men,median age:45 years;interquartile range:39-50 years)enrolled in an OTP between October 2015 and September 2017.Patients were followed until September 2019.Data on sociodemographics,substance use,HCV infection,human immunodeficiency virus(HIV)infection and laboratory parameters were collected at entry.We analyzed medical records to evaluate HCV treatment.Kaplan-Meier methods and Cox regression models were used to analyze the DAA treatment uptake and to identify treatment predictors.RESULTS Prevalence of HCV and HIV infection was 70%and 34%,respectively.Among anti-HCV-positive(n=336)patients,47.2%,41.3%,and 31.9%used alcohol,cannabis,and cocaine,respectively.HCV-RNA tests were positive in 233(69.3%)patients.Twentyeight patients(8.3%)cleared the infection,and 59/308(19.1%)had received interferon-based treatment regimens before 2015.Among 249 patients eligible,111(44.6%)received DAAs.Treatment rates significantly increased over time from 7.8/100 person-years(p-y)(95%CI:5.0-12.3)in 2015 to 18.9/100 p-y(95%CI:11.7-30.3)in 2019.In a multivariate analysis,patients with HIV co-infection were twice as likely to receive DAAs(HR=1.94,95%CI:1.21-3.12)than patients with HCV mono-infection.Current drug use was an independent risk factor for not receiving treatment against infection(HR=0.48,95%CI:0.29-0.80).CONCLUSION HCV treatment is evolving in patients with HCV-HIV co-infection.Ongoing drug use while in an OTP might negatively impact the readiness to treat infection.

Interferon-free treatments in patients with hepatitis C genotype 1-4 infections in a real-world setting

AIM To investigated the real-world effectiveness and safety of various regimens of interferon-free treatments in patients infected with hepatitis C virus(HCV).METHODS We performed an observational study to analyze different antiviral treatments administered to 462 HCV-infected patients, of which 56.7% had liver cirrhosis. HCV RNA after 4 wk of treatment and at 12 wk after treatment sustained virologic response(SVR) as well as serious adverse events(SAEs) was analyzed first for the whole cohort and then separately in patients who met or did not meet the inclusion criteria of a clinical trial(CT-met and CT-unmet, respectively).RESULTS The most frequently prescribed treatment was simeprevir/sofosbuvir(36.4%), followed by sofosbuvir/ledipasvir(24.9%) and ombitasvir/paritaprevir/ritonavir(r)/dasabuvir(19.9%). Ribavirin(RBV) was administered in 198 patients(42.9%). SVRs occurred in 437/462 patients(94.6%). The SVRs ranged between 93.3% and 100% for genotypes 1-4. SVRs were achieved in 96.2% patients in the CTmet group vs 91.9% patients in the CT-unmet group(P = 0.049). Undetectable HCV RNA at week 4 occurred in 72.9% of the patients. In the univariate analysis, the factors associated with SVRs were lower liver stiffness, absence of cirrhosis, higher platelet count, higher albumin levels, no RBV dose reduction, undetectable HCV RNA at week 4 and CT-met group. In the multivariate analysis, only albumin was an independent predictor of treatment failure(P = 0.04). Eleven patients(2.4%) developed SAEs; 5.2% and 0.7% of the patients in the CT-unmet and CT-met groups, respectively(P = 0.003).CONCLUSION A high proportion of patients with HCV infection achieved SVRs. For patients who did not meet the CT criteria, treatment regimens must be optimized.

Global prevalence of occult hepatitis C virus: A systematic review and meta-analysis

BACKGROUND Occult hepatitis C infection(OCI)is characterized by the presence of hepatitis C virus(HCV)RNA in the liver,peripheral blood mononuclear cells(PBMC)and/or ultracentrifuged serum in the absence of detectable HCV-RNA in serum.OCI has been described in several categories of populations including hemodialysis patients,patients with a sustained virological response,immunocompromised individuals,patients with abnormal hepatic function,and apparently healthy subjects.AIM To highlight the global prevalence of OCI.METHODS We performed a systematic and comprehensive literature search in the following 4 electronic databases PubMed,EMBASE,Global Index Medicus,and Web of Science up to 6th May 2021 to retrieve relevant studies published in the field.Included studies were unrestricted population categories with known RNA status in serum,PBMC,liver tissue and/or ultracentrifuged serum.Data were extracted independently by each author and the Hoy et al tool was used to assess the quality of the included studies.We used the random-effect meta-analysis model to estimate the proportions of OCI and their 95%confidence intervals(95%CI).The Cochran's Q-test and the I2 test statistics were used to assess heterogeneity between studies.Funnel plot and Egger test were used to examine publication bias.R software version 4.1.0 was used for all analyses.RESULTS The electronic search resulted in 3950 articles.We obtained 102 prevalence data from 85 included studies.The pooled prevalence of seronegative OCI was estimated to be 9.61%(95%CI:6.84-12.73)with substantial heterogeneity[I^(2)=94.7%(95%CI:93.8%-95.4%),P<0.0001].Seropositive OCI prevalence was estimated to be 13.39%(95%CI:7.85-19.99)with substantial heterogeneity[I^(2)=93.0%(90.8%-94.7%)].Higher seronegative OCI prevalence was found in Southern Europe and Northern Africa,and in patients with abnormal liver function,hematological disorders,and kidney diseases.Higher seropositive OCI prevalence was found in Southern Europe,Northern America,and Northern Africa.CONCLUSION In conclusion,in the present study,it appears that the burden of OCI is high and variable across the different regions and population categories.Further studies on OCI are needed to assess the transmissibility,clinical significance,long-term outcome,and need for treatment.

Pregnancy and fetal outcomes of chronic hepatitis C mothers with viremia in China

BACKGROUND Data that assess maternal and infant outcomes in hepatitis C virus(HCV)-infected mothers are limited.AIM To investigate the frequency of complications and the associated risk factors.METHODS We performed a cohort study to compare pregnancy and fetal outcomes of HCVviremic mothers with those of healthy mothers.Risk factors were analyzed with logistic regression.RESULTS Among 112 consecutive HCV antibody-positive mothers screened,we enrolled 79 viremic mothers.We randomly selected 115 healthy mothers from the birth registry as the control.Compared to healthy mothers,HCV mothers had a significantly higher frequency of anemia[2.6%(3/115)vs 19.0%(15/79),P<0.001]during pregnancy,medical conditions that required caesarian section[27.8%(32/115)vs 48.1%(38/79),P=0.004],and nuchal cords[9.6%(11/115)vs 34.2%(27/79),P<0.001].In addition,the mean neonatal weight in the HCV group was significantly lower(3278.3±462.0 vs 3105.1±459.4 gms;P=0.006),and the mean head circumference was smaller(33.3±0.6 vs 33.1±0.7 cm;P=0.03).In a multivariate model,HCV-infected mothers were more likely to suffer anemia[adjusted odds ratio(OR):18.1,95%confidence interval(CI):4.3-76.6],require caesarian sections(adjusted OR:2.6,95%CI:1.4-4.9),and have nuchal cords(adjusted OR:5.6,95%CI:2.4-13.0).Their neonates were also more likely to have smaller head circumferences(adjusted OR:2.1,95%CI:1.1-4.3)and lower birth weights than the average(≤3250 gms)with an adjusted OR of 2.2(95%CI:1.2-4.0).The vertical transmission rate was 1%in HCV-infected mothers.CONCLUSION Maternal HCV infections may associate with pregnancy and obstetric complications.We demonstrated a previously unreported association between maternal HCV viremia and a smaller neonatal head circumference,suggesting fetal growth restriction.

The association between the genetic polymorphisms of LMP2/LMP7 and the outcomes of HCV infection among drug users

Objective：To investigate a possible association of LMP2/LMP7 genes with chronic hepatitis C virus（HCV） infection,and to assess whether LMP2/LMP7 genes could influence the outcomes of HCV infection among drug users.Methods：Genomic DNAs of 362 anti-HCV sero-positive drug users and 225 control drug users were extracted from the peripheral blood leukocytes.The sero-positive patients were divided into those who had persistent infection and those who had spontaneously cleared the infection.Polymorphisms of LMP genes were determined by PCR combined with restriction fragment length polymorphism（RFLP）.Results：The distribution of LMP2 genotypes among the control,persistent infection and spontaneous clearance groups were not different.However,the LMP7 codon 145 Gln/Lys,Lys/Lys,and Gln/Lys＋Lys/Lys genotypes were found significantly more frequent in the persistent infection group than in control group（OR=1.75,95%CI=1.06～2.90;OR=3.16,95%CI=1.23-8.12;OR=1.94,95%CI=1.21-3.12,respectively）.Similarly,the frequencies of the codon 145 Gln/Lys,Lys/Lys,and Gln/Lys＋Lys/Lys genotypes were found significantly more frequent in the persistent infection group than in the spontaneous clearance group（OR=1.64,95%CI=1.04-2.57;OR=2.40,95%CI=1.09-5.28;OR=1.76,95%CI=1.152.69,respectively）.Stratified analysis indicated that combined genotype Gln/Lys＋Lys/Lys of the LMP7 gene was related to an increasing susceptibility to HCV infection（OR=1.91,95%CI=1.02-3.55;OR=2.19,95%CI=1.243.89;OR=1.91,95%CI=1.05-3.48,OR=2.86,95%CI=1.41-5.78,respectively）and the risk of persistent HCV infection（OR=1.94,95%CI=1.12-3.34;OR=2.02,95%CI=1.21-3.38;OR=1.78,95%CI=1.12-2.85,OR=2.23,95%CI=1.09-4.58,respectively）among30-year-old,males,the injection drug user（IDU）subjects and/or the shorter duration drug users（≤5 y）.Conclusion：These results suggest that polymorphism of the LMP7 gene may have an influence on the outcomes of HCV infection,and is one of the factors accounting for the genetic susceptibility to HCV infection among drug users.

Transient elastography: A non-invasive tool for assessing liver fibrosis in HIV/HCV patients

AIM: To assess the prevalence of advanced liver fibrosis (ALF) in human immunodeficiency virus (HIV), hepatitis C virus (HCV) and HIV/HCV patients using transient elastography, and to identify factors associated with ALF. METHODS: Between September 2008 and October 2009, 71 HIV mono-infected, 57 HIV/HCV co-infected and 53 HCV mono-infected patients on regular follow-up at our Center were enrolled in this study. Alcohol intake, the main parameters of liver function, presence of HCV-RNA, HIV-RNA, duration of highly active anti-retroviraltherapy (HAART) and CD4 cell count were recorded. ALF was defined as liver stiffness (LS) ≥ 9.5 kPa. To estimate liver fibrosis (LF) a further 2 reliable biochemical scores, aspartate aminotransferase platelet ratio index (APRI) and FIB-4, were also used. RESULTS: LS values of co-infected patients were higher than in either HIV or HCV mono-infected patients (χ 2M H = 4, P < 0.04). In fact, LS ≥ 9.5 was significantly higher in co-infected than in HIV and HCV mono-infected pa-tients (χ 2 = 5, P < 0.03). Also APRI and the FIB-4 index showed more LF in co-infected than in HIV mono-infect-ed patients (P < 0.0001), but not in HCV mono-infected patients. In HIV?HCV co-infected patients, the extent of LS was significantly associated with alcohol intake (P < 0.04) and lower CD4+ cell count (P < 0.02). In HCV pa-tients, LS was correlated with alcohol intake (P < 0.001) and cholesterol levels (P < 0.03). Body mass index, dia-betes, HCV-and HIV-viremia were not significantly cor-related with LS. In addition, 20% of co-infected patients had virologically unsuccessful HAART; in 50% compliance was low, CD4+ levels were < 400 cells/mm 3 and LS was > 9.5 kPa. There was no significant correlation between extent of LF and HAART exposure or duration of HAART exposure, in particular with specific dideoxynucleoside analogues. CONCLUSION: ALF was more frequent in co-infected than mono-infected patients. This result correlated with lower CD4 levels. Protective immunological effects of HAART on LF progression outweigh its hepatotoxic effects.

The impact of direct-acting antivirals on hepatitis C associated hepatocellular carcinoma

The increased incidence of hepatocellular carcinoma(HCC)in the last several decades in the United States and worldwide has partly resulted from an increase in hepatitis C virus(HCV)infection.HCV carcinogenesis is speculated to be indirectly related to multiple steps from inflammation to fibrosis and advanced fibrosis/cirrhosis over 20 or more years.However,the direct carcinogenic potential from HCV may explain HCC occurring in non-cirrhotic HCV patients.Highly potent direct-acting antivirals(DAAs)in recent years have changed hepatitis C treatment significantly and have resulted in the sustained virologic response(SVR)rate exceeding 90%.Although initial reports concerned the increase in de novo and recurrent HCC associated with DAAs,more recent studies showed that DAA-induced SVR on the contrary reduced risk of HCV-associated HCC without increasing its recurrence.The International Consortium of Hepatitis C Therapeutic Registry and Research Network(HCV-TARGET)database and other resources of HCV patients treated with DAA collectively in the near future most likely will be able to show definitive evidence on the risk of HCC occurrence and recurrence after DAA with SVR.The long-term risk of HCC in chronic hepatitis C patients with advanced fibrosis or cirrhosis remains high after DAAs with SVR.Thus,HCC surveillance on this sub-group of patients is important for early detection and intervention of HCC.

NKT cells in liver diseases

Natural killer T cells are innate-like and tissue-resident lymphocytes, which recognize lipid antigens and are enriched in the liver. Natural killer T cells play important roles in infections, tumors, autoimmune diseases, and metabolic diseases. In this study, we summarize recent findings on biology of natural killer T cells and their roles in hepatitis B virus and hepatitis C virus infection, autoimmune liver diseases, alcoholic liver disease, nonalcoholic fatty liver disease, and hepatocellular carcinoma. Controversial results from previous studies are discussed, and indicate the dynamic alteration in the role of natural killer T cells during the progression of liver diseases, which might be caused by changes in natural killer T subsets, factors skewing cytokine responses, and intercellular crosstalk between natural killer T cells and CDld-expressing cells or bystander cells.

Hepatocellular carcinoma in HCV - liver cirrhosis before and after successful DAA treatment

Chronic hepatitis C virus(HCV)infection is a major cause of liver cirrhosis and hepatocellular carcinoma(HCC)worldwide.The recent advancement of direct-acting Antiviral Agents(DAAs)in hepatitis C therapy,resulted in sustained virological response rates of over 90%in treated patients in different stages of liver fibrosis.The efficacy of DAAs treatment has also been confirmed in real-life cohorts that include subjects with decompensated cirrhosis and therefore seems a promising step to a significant reduction in the recurrence of HCC in patients who achieved complete destruction of the HCC nodules by local therapy.We present a 72-year old patient with HCV-related liver cirrhosis who successfully responded to DAAs treatment after complete destruction of an early HCC nodule.