A pilot study of daily subcutaneous interleukin-10 in patients with chronic hepatitis C infection.

The Th1/Th2 cytokine balance is important in persistence of infection and liver injury in chronic hepatitis C. The aim of this study was to administer the anti-inflammatory cytokine, recombinant human interleukin-10 (rHuIL-10), for 28 days in patients with chronic hepatitis C and to assess the safety and measure the effect on alanine aminotransferase (ALT, a marker of hepatic inflammation) levels and serum hepatitis C virus (HCV) RNA values. Three treatment-naive and 13 interferon (IFN) nonresponder patients (total 16 patients) with compensated chronic HCV infection were enrolled in this study. Patients were randomized to receive rHuIL-10 at a dose of 4 or 8 microg/kg/day as a single daily subcutaneous injection for 28 days. ALT values and serum HCV RNA were measured at days 0, 1, 3, 8, 15, 22, and 28 during therapy and at follow-up 2 and 4 weeks after cessation of the 4-week treatment period. ALT values normalized in 9 of 16 patients during therapy and remained normal until the end of treatment in 8 patients. The decreases in ALT values occurred in both the 4 microg and 8 microg dosage groups and were seen in both IFN naive and nonresponder patients. Mean ALT values fell significantly during the study period but usually returned to pretreatment levels by the end of the 4-week follow-up period (p < 0.05). HCV RNA concentrations did not vary significantly during or after therapy. (No patient had either an increase or a decrease in HCV RNA levels of > or =1.5 log during the study.) The drug was well tolerated, with no adverse symptoms noted. Platelet counts fell transiently to 73,000 and 63,000 in 2 patients. No other toxicity was observed, and no patients discontinued therapy. In chronic hepatitis C, short-term therapy with IL-10 was well tolerated and caused transient normalization of ALT values in 50% of patients, which returned to pretreatment levels on cessation of treatment. There were no significant changes observed in serum HCV RNA concentrations during the study. These immunomodulatory effects are similar to those observed with ribavirin monotherapy in chronic hepatitis C. Further study of rHuIL-10 alone or in combination with antiviral agents in chronic hepatitis C is warranted.     

Impaired filterability of erythrocytes from patients with chronic hepatitis C and effects of eicosapentaenoic acid on the filterability

Although erythrocyte filterability plays a key role in microcirculation, it is unknown whether the filterability of erythrocytes from patients with chronic hepatitis C (CH-C) is impaired. This study aimed to investigate erythrocyte filterability in CH-C patients in relation to medical treatment. The mean erythrocyte filterability (%) for all 24 patients with CH-C (69.2 +/- 10.8%) was significantly lower than that for 5 normal controls (80.5 +/- 1.7%, P < 0.03). In 8 patients, the combination therapy of ribavirin (RBV) and interferon improved liver function but caused anemia. The filterability after treatment (57.8 +/- 12.8%) was lower than that before treatment (70.8 +/- 9.7%, P < 0.05). Decreased filterability showed no correlation with the mean corpuscular volume or mean corpuscular Hb concentration during treatment, suggesting that the decrease in filterability mainly arises from changes in erythrocyte membrane properties. We investigated the protective effects of eicosapentaenoic acid (EPA) on the RBV-induced anemia. Filterability in 7 responders was markedly improved from 68.4 +/- 4.6% to 77.4 +/- 2.4% (P < 0.001), but not in 3 nonresponders. In the responders, the progression of anemia was restrained. In conclusion, we found an obvious impairment of the filterability of erythrocytes from CH-C patients, further impairment of the filterability induced by oxidative membrane damage caused by RBV leading to hemolytic anemia, and amelioration of the filterability caused by the antioxidative effects of EPA.     

Add‐on therapy of pitavastatin and eicosapentaenoic acid improves outcome of peginterferon plus ribavirin treatment for chronic hepatitis C

Despite the use of pegylated‐interferon (peg‐IFN) plus ribavirin combination therapy, many patients infected with hepatitis C virus (HCV)‐1b remain HCV‐positive. To determine whether addition of pitavastatin and eicosapentaenoic acid (EPA) is beneficial, the “add‐on” therapy option (add‐on group) was compared retrospectively with unmodified peg‐IFN/ribavirin therapy (standard group). Association of host‐ or virus‐related factors with sustained virological response was assessed. In HCV replicon cells, the effects of pitavastatin and/or EPA on HCV replication and expression of innate‐immunity‐ and lipid‐metabolism‐associated genes were investigated. In patients infected with HCV‐1b, sustained virological response rates were significantly higher in the add‐on than standard group. In both groups, sustained virological response rates were significantly higher in patients with genotype TT of IL‐28B (rs8099917) than in those with non‐TT genotype. Among the patients with non‐TT genotype, sustained virological response rates were markedly higher in the add‐on than standard group. By multivariate analysis, genome variation of IL28B but not add‐on therapy remained as a predictive factor of sustained virological response. In replicon cells, pitavastatin and EPA suppressed HCV replication. Activation of innate immunity was obvious in pitavastatin‐treated cells and EPA suppressed the expression of sterol regulatory element binding protein‐1c and low‐density lipoprotein receptor. Addition of pitavastatin and EPA to peg‐IFN/ribavirin treatment improved sustained virological response in patients infected with HCV‐1b. Genotype variation of IL‐28B is a strong predictive factor in add‐on therapy. J. Med. Virol. 85:250–260, 2013. © 2012 Wiley Periodicals, Inc.     

Iron overload in patients with chronic hepatitis C: A clinicopathologic study

Recent studies suggest that increased hepatic iron may impair the response to interferon therapy in patients with chronic hepatitis C. We reviewed the records and liver biopsies of 72 patients with chronic hepatitis C to determine the prevalence of iron overload and to evaluate whether there is a correlation between serum and hepatic iron concentrations and activity of liver disease. Patients with other causes of liver disease or iron overload were excluded. Necroinflammatory activity and fibrosis were evaluated using modified Knodell score. Hepatic iron was assessed using Brissot's grading system. Increased serum iron and ferritin levels were found in 29% and 43% patients, respectively. Hepatic iron grades 0, I, II, III, and IV were present in 37%, 35%, 25%, 3%, and 0% of patients, respectively. A significant correlation was found between hepatic iron grade and serum ferritin (P = .0001). There was no correlation between hepatic iron grade and histological activity index or fibrosis score. In summary, we found a high proportion of patients with chronic hepatitis C had mild to moderate increase in hepatic iron content even when patients with alcoholism and recurrent transfusions were excluded. However, very few patients had severely increased iron load.     

A prospective study of neuropsychiatric symptoms associated with interferon-alpha-2b and ribavirin therapy for patients with chronic hepatitis C

The purpose of this study was to systematically describe the neuropsychiatric side effects of treatment with interferon-alpha-2b (INF-alpha) and ribavirin in patients with chronic hepatitis C as well as to compare different instruments used to measure these side effects. Fifty-five patients with chronic hepatitis C were prospectively followed for 24 weeks and assessed with seven neuropsychiatric symptom measures and one quality of life scale. Of 42 patients treated with INF-alpha and ribavirin, 11 (26%) were receiving psychiatric treatment at baseline. They scored higher on all rating scales at baseline and became more symptomatic during treatment. Of the 31 patients (74%) not in psychiatric care at baseline, 15 (48%) required treatment for neuropsychiatric symptoms, and seven (23%) met criteria for major depression during INF-alpha therapy. The control group of 13 untreated subjects showed little change over the 24-week period. All symptom scales were highly intercorrelated, suggesting that use of one is sufficient for monitoring symptoms.     

Pernicious anemia during IFN-alpha treatment for chronic hepatitis C.

Some latent diseases, such as immune disorders, can appear during interferon-alpha (IFN-alpha) therapy. These disorders are difficult to predict because of their low prevalence in the general population. We describe a case of pernicious anemia (PA) in a patient affected by chronic hepatitis C and macrocytosis during IFN-alpha therapy. Hemoglobin (Hb) concentration reached 7.3 g/dl. Anti-intrinsic factor (IF) antibodies were present, but not antiparietal cell antibodies (APCA). Suspension of IFN-alpha and administration of vitamin B(12) resulted in normal Hb concentrations. This case is the first instance of early PA (at the second month of IFN therapy) in a patient affected by chronic hepatitis C. The only other case of PA in a patient affected by hepatitis C virus (HCV) infection occurred during the second year of maintenance IFN therapy. We recommend that particular attention be paid to such clinical and laboratory conditions as macrocytosis in administering IFN-alpha therapy for chronic hepatitis C.     

慢性丙型肝炎病毒感染者血清自身抗体检测的临床研究

目的 观察丙型肝炎病毒感染者血清中自身抗体的检测结果,探讨自身免疫在丙型肝炎病毒感染中的意义.方法 采用回顾性分析方法,对226例慢性丙型肝炎病毒感染者和137例慢性乙型肝炎患者血清进行ANA、anti-AMA、anti-Ro-52等自身抗体的检测,并探讨HCV-RNA含量、生化指标、年龄及性别、干扰素治疗后应答等与自身抗体变化的关系.结果 丙型肝炎感染者226例中有78例出现自身抗体阳性,检出率为34.5%（78/226）;明显高于慢性乙肝组的7.3%（10/137）（χ^2=34.396,P＜0.05）,其中ANA阳性69例,占30.5%.抗核抗体以低滴度为主.另外还检测到anti-AMA、anti-Ro-52等自身抗体;150例丙肝病毒复制指标HCV-RNA（＋）的患者,自身抗体检出率为40.7%,76例丙肝病毒复制指标HCV-RNA（-）的患者,自身抗体检出率为22.4%,两者总检出率差别有统计学意义（χ^2=7.473,P＜0.05）;自身抗体阴性、阳性者之间ALT、AST、TBIL数值分别为（65.1±24.4）U/L、（47.4±22.7）U/L、（17.2±8.2）μmol/L和（132.2±49.3）U/L、（100.7±35.2）U/L、（35.5±14.7）μmol/L,差异有统计学意义（t值分别为16.012,14.843,和9.000,均P＜0.05）;丙型肝炎自身抗体的检出率与年龄有关,而与性别无关;丙型肝炎患者经干扰素抗病毒治疗后,虽然自身抗体阳性组干扰素应答率73.9%（17/23）高于自身抗体阴性组的54.2%（26/48）,但差异无统计学意义（χ2=0.975,P＞0.05）.结论 丙型肝炎病毒感染可诱发机体自身免疫反应,产生多种自身抗体,尤其HCV-RNA阳性患者更为突出,自身抗体检出率与年龄明显有关,检测自身抗体对丙型肝炎的诊断治疗具有指导意义.     

Retreatment of patients with chronic hepatitis C

Impressive gains have been made in the treatment of chronic hepatitis C virus (HCV) infection during the past decade. As the treatment for chronic hepatic C has improved, the question has arisen as to whether patients who failed previous HCV treatment regimens should be retreated. Several recent studies involving retreatment have established that a small but significant increase in sustained virologic response (SVR) resulted when nonresponders to prior interferon monotherapy were retreated with interferon and ribavirin. Given the superior results observed with pegylated interferon and ribavirin in the treatment of naive population, it is now appropriate to consider whether retreating previous relapsers and nonreponders, particularly nonresponders to conventional interferon and ribavirin, will be effective. Such treatment would be most advantageous for patients with advanced fibrosis or cirrhosis who are at the highest risk of developing complications of advanced liver disease, including hepatocellular carcinoma. Namely, there is no doubt that interferon-based regimens may reduce hepatic inflammation in the abscence of SVR. For this reason, patients with advanced liver disease receive histologic benefit from therapy in addition to the potential benefit of viral eradication, because progression of fibrosis to cirrhosis is a function of hepatic inflammation. A number of investigator-initiated clinical trials have been started to study response in prior nonresponders and relapsers retreated with pegylated interferon and ribavirin. From the data obtained from these trials, so far is clear that combined therapy can clear HCV-RNA in a significant number of prior nonresponders (25% to 40%) while on treatment, yet the rate of SVR appears to be by far lower (20%). Better results have been reported with relapsers to prior combination therapy. Moreover, preliminary results from the HALT-C trial have shown that maintenance therapy with pegylated interferon and ribavirin can significantly delay fibrosis progression and reduce the risk of hepatic decompensation and hepatocellular carcinoma and the need of liver transplantation. According to these results, many authors have proposed a course of combination therapy with pegylated interferon and ribavirin for relapsers and nonresponders with advanced fibrosis and cirrhosis.     

Efficacy of long-term interferon therapy in chronic hepatitis B patients with HBV genotype C

Infection with Hepatitis B virus (HBV) genotype C predominates in Japan. We analyzed the efficacy of interferon (IFN) alpha or beta in the treatment of chronic hepatitis B patients with HBV genotype C and the clinical predictors for therapeutic response. Forty-three genotype C-infected, chronic hepatitis B e antigen (HBeAg)-positive patients (32 men and 11 women with a mean age of 35.6+/-10.1 years) who had been treated with IFN therapy were retrospectively studied. The patients were classified into two treatment groups. Short-term therapy group was administered a 5-6 MU dose three times weekly for 4 weeks, and the long-term therapy group for 24 weeks. At the end of the follow-up period, 4 (15%) of 27 short-term therapy group patients and 6 (38%) of 16 long-term therapy group patients had normalized serum ALT levels and seroconversion of HBeAg to anti-HBe (p=0.137). Multivariate analysis for parameters most important for the efficacy of IFN therapy was performed using Cox proportional hazard models in order to investigate the association between baseline characteristics of patients and the response to IFN treatment. As a result, the p-values of IFN treatment group and sex were <0.05, and both factors can be recognized as independent significant factors (relative risk, 2.93 and 2.53; p=0.027 and 0.040, respectively). Furthermore, the cumulative rates of seroconversion of HBeAg to anti-HBe analyzed by the Kaplan-Meier method was significantly higher in the female group (p=0.015) and in the long-term IFN therapy group (p=0.0046). In summary, long-term IFN therapy may be more effective than short-term IFN therapy for patients with chronic HBV genotype C infection.     

Longitudinal study of hepatitis C viremia in chronic hepatitis C

Serial serum samples from 20 untreated patients with chronic hepatitis C virus (HCV) infection were tested for HCV RNA by a nested polymerase chain reaction assay using primers from the highly conserved 5′ noncoding region to determine the relationship between hepatitis C viremia and the activity of liver disease during the natural course of chronic HCV infection. Semiquantitation of serum HCV RNA level was achieved by testing serial 10-fold dilutions of RNA extracts to determine the end-point titer. All the patients were HCV RNA positive at presentation. There was a poor correlation between the initial HCV RNA titer and serum transaminase levels. All patients except one were persistently HCV RNA positive during a follow-up period of 1.5-15 years, although 17 (85%) had periods of normal or near-normal transaminase levels. There was no correlation between changes in the serum transaminase levels and HCV RNA titer. Patients with chronic HCV infection have persistent viremia despite fluctuations in ALT levels.     

Hepatitis C virus RNA in patients with chronic hepatitis C

Reviews recent data on the detection of genome and replicative HCV RNA in patients with chronic hepatitis C by PCR and in situ hybridization. Discusses the results of HCV RNA detection in the liver, lymphocytes, serum, and other organs and tissues and notes the relationship between the incidence of RNA and activity of the pathological process. Analyzes the results of HCV RNA detection after IFN treatment. Discusses the role of HCV RNA in the pathogenesis of hepatitis C.     

重组干扰素对慢性丙型肝炎抗病毒疗效5年随访观察

目的观察重组干扰素α－２ａ，α－２ｂ抗丙型肝炎病毒（ＨＣＶ）的近、远期疗效。方法重组干扰素α－２ａ治疗组７０例，重组干扰素α－２ｂ４６例对照组２８例，治疗后随访５年。结果治疗结束时，ＨＣＶＲＮＡ阴转率和血清ＡＬＴ复常率α－２ａ组分别为６７１４％和７０００％，α－２ｂ组分别为６９５６％和７１７３％，随访５年后，α－２ａ组ＨＣＶＲＮＡ阴转率和血清ＡＬＴ复常率分别为３５７１％和４７１４％，α－２ｂ组分别为３９１３％和５２１７％，均显著高于对照组（Ｐ＜００１和Ｐ＜００５）。基因分型以ＨＣＶⅠ组感染为主（７５８６％），干扰素对ＨＣＶⅡ组感染的疗效优于ＨＣＶⅠ组。结论重组干扰素α－２ａ与α－２ｂ均为有效的抗丙型肝炎病毒药物，慢性丙型肝炎患者干扰素治疗的早期疗效较好。ＨＣＶ基因型有预测干扰素疗效的意义     

Dynamics of serum hepatitis C virus load and quasispecies complexity during antiviral therapy in patients with chronic hepatitis C.

BACKGROUND: Hepatitis C virus (HCV) infection is a dynamic process during which viral genetic variants continuously develop as a result of the virus adaptation to the host's immune system. The level of viremia and the complexity of the hypervariable region 1 (HVR 1) quasispecies of hepatitis C virus during antiviral therapy reflect the dynamic balance between the viral and host components in response to therapy. OBJECTIVE: The aim of the study was to evaluate the dynamics of HCV viremia and the complexity of the HVR 1 quasispecies during the induction phase of a triple combination therapy regimen in nonresponders to earlier anti-HCV treatment. STUDY DESIGN: Ten patients with chronic hepatitis C undergoing antiviral combination therapy with interferon-alpha, ribavirin, and amantadine were studied. The serum HCV RNA level was monitored by a quantitative RT-PCR assay up to 3 months after start of treatment. The HVR 1 quasispecies complexity was analysed by an "in house" nested RT-PCR mediated single-strand conformation polymorphism (SSCP) assay. RESULTS: Baseline serum HCV RNA levels ranged from 1.94x10(6) to 5.53x10(6) copies/ml. In all patients, HCV subtype 1b was found. At the start of therapy, the SSCP assay revealed a high complexity pattern (at least six SSCP bands) in all patients. None of the patients responded within 4 weeks of treatment, however, the serum HCV RNA level decreased by one to two logs in eight patients. At week 4 after start of treatment, there was a decrease of SSCP bands in five patients. In four patients, SSCP bands remained unchanged and in one patient SSCP bands increased. At month 3 after start of treatment, serum HCV RNA was not detectable in one patient. CONCLUSION: Because of the low number of patients involved in this study, prediction of therapeutical success based on the quasispecies complexity was not possible. Larger studies are urgently needed.     

A replicon-based bioassay for the measurement of interferons in patients with chronic hepatitis C

Overall treatment results of chronic hepatitis C have improved markedly with the introduction of pegylated interferon-alpha (PEG–IFN-) and ribavirin combination therapy. However, cure rates in the most common genotype 1 infection are still unsatisfactory. IFN- dose–response studies on viral kinetics suggest that inadequate dosing might be a key factor but drug levels have hardly been tested, which is in part due to difficulties in measuring this cytokine in patient samples. We have shown recently that hepatitis C virus (HCV) replicons are highly sensitive to IFN-. In this report we tested whether the replicon system could be used as a sensitive bioassay to determine the amount of biologically active IFN- in serum or heparinized plasma of patients under therapy. To facilitate the measurements, a stably replicating subgenomic HCV RNA was developed that carries the gene encoding the firefly luciferase. Dose response studies with IFN- demonstrate that the amount of expressed luciferase directly correlates with the level of HCV replication. By using this cell-based assay, serum samples of HCV patients treated with different types and doses of IFN- were analyzed in parallel to IFN- standards made by serial dilutions of the same type of IFN- the patient was treated with. Based on nonlinear logistic models serum concentrations corresponding to 1.3–19 U/ml were determined in patients under standard or high dose IFN- therapy, and from 3.8 to 4.1 ng/ml in patients treated with PEG IFN-. In conclusion, the HCV-replicon based bioassay allows determining the levels of biologically active IFN- in serum and heparinized plasma of patients under treatment.     

Hepatitis C virus amino acid sequence diversity correlates with the outcome of combined interferon/ribavirin therapy in Chinese patients with chronic hepatitis C

Evidence has shown that the p7, NS2 and NS3 genes affect the outcome of pegylated-IFN-α/ribavirin (PEG-IFN/RBV) combination therapy in different populations with HCV infections. Here, we test the hypothesis that diversity in the p7, NS2 and NS3 genes influences the probability of obtaining either a sustained (SVR) or non-sustained (non-SVR) viral response in Chinese patients with genotype 1b chronic hepatitis C. There were significantly more unique variations in the p7, NS2 and NS3 genes in the sequences from SVR than non-SVR patients. Inter-patient variations related to treatment outcome in NS3 were concentrated in the protease domain. There were no significant differences in the frequency of variations in the core, E1 and E2 proteins between the groups. In conclusion, increased amino acid sequence diversity in the p7, NS2 and NS3 genes is associated with an SVR to PEG-IFN/RBV therapy in Chinese patients with genotype 1b chronic hepatitis C.     

Novel approaches for therapy of chronic hepatitis C.

Currently available anti-HCV therapy is effective in only half of the patients and limited by side effects that often necessitate discontinuation. Therefore, new treatment strategies are being developed including (i) the optimization of current regimens, (ii) the use of additional agents working via novel mechanisms, and (iii) anti-fibrotic strategies. Many new antiviral compounds are now being studied in preclinical and clinical trials. This review will focus on drugs that have already entered the stage of phase 2 or phase 3 studies.     

Prolonged-interferon therapy reduces hepatocarcinogenesis in aged-patients with chronic hepatitis C

The aim of this study was to elucidate the reduction of hepatocarcinogenesis by prolonged interferon (IFN) monotherapy in aged chronic hepatitis C patients. Inclusion criteria were biopsy-proven chronic hepatitis or liver cirrhosis, 60 years and over, elevated serum aminotransferase and positive hepatitis C virus (HCV)-RNA. One hundred and twenty patients satisfied the above criteria were treated with natural IFN-alpha (dose: 3 million unit (MU), two or three times weekly for 0.5-15.5 years, mean 2.47 years) (IFN group). Another 240 patients treated with herbal medicines excluding IFN were selected as control (no-IFN group). The patients not treated with IFN were matched 2:1 with IFN group patients for sex and age. The clinical and biological differences were compared after treatment with the IFN group and the untreated group. Serum alpha-fetoprotein (AFP) level decreased with statistical significance after initiation of treatment with IFN compared to no treatment. The 5- and 10-year cumulative rates of hepatocellular carcinoma (HCC) were 5.9 and 13.7%, and 17.1 and 32.8%, for the IFN and untreated group, respectively. HCC development occurred when histologic staging was advanced, and IFN was not given, the AFP level after treatment was >10 ng/ml. Cox regression analysis indicated that the relative risk of HCC in patients in the IFN group was 0.3 times of that in the untreated patients. The relative risk rate for HCC in severe fibrosis was 3.9 compared with mild or moderate fibrosis. In conclusion, long-term IFN therapy for aged patients with chronic HCV infection is effective in decreasing the serum AFP level and preventing hepatocarcinogenesis.     

Management of patients with chronic hepatitis C infection

Abstract Chronic infection with hepatitis C virus (HCV) is a leading cause of liver-related morbidity and mortality throughout the world. Although reliable figures regarding the global prevalence of HCV infection are wanting, it is likely that HCV prevalence will continue to increase. Injection drug use is the most important source of HCV transmission in the developed world, while unsafe therapeutic injection is an important source of transmission in developing nations. The majority of exposed individuals become chronically infected, of whom 50% develop chronic liver injury. Cirrhosis and hepatocellular carcinoma can arise in those chronically infected over a mean of 20–30 years. Despite this high prevalence and morbidity, recommendations regarding who to screen by antibody testing remain disparate. Quantitative measurement of HCV RNA and HCV genotyping is useful in predicting response to antiviral therapy. Noninvasive methods of detecting liver injury, such as serologic batteries, have not been as informative or predictable as liver biopsy. The current pharmacologic standard of care for chronic HCV infection is the combination of subcutaneous peginterferon and oral ribavirin, which yields sustained virologic response in 54%–56%. Higher rates of SVR are seen in those patients who are infected with HCV genotypes 2 and 3. As intravenous drug use remains the most important source of HCV transmission in the US and Europe, education within this group is an important preventive tool.