血清乙肝病毒大蛋白与乙肝前S1抗原联合检测在HBeAg阴性乙肝患者诊断中的意义

目的 探讨乙型肝炎e抗原阴性[HBeAg（－）]乙肝患者血清乙肝病毒大蛋白（HBV-LP）和乙肝前S1抗原（PreS1-Ag）联合检测的临床意义。方法 采用酶联免疫吸附（ELISA）法测定300例慢性乙肝患者的血清HBV-LP和PreS1-Ag浓度;实时荧光定量PCR（qRT-PCR）法检测血清HBV-DNA表达量;比较不同HBV-M模式下HBV-LP、PreS1-Ag与HBV-DNA的阳性检出率;分析以HBV-DNA表达量作为HBV感染及复制的金标准时,血清HBV-LP和PreS1-Ag单独检测及联合检测对HBeAg阴性乙肝患者的阳性预测值和阴性预测值。结果 （1）115例HBeAg（+）血清中,HBV-LP和PreS1-Ag的阳性率均与HBV-DNA阳性率差异无统计学意义（Ps＞0.05）;120例HBeAg（－）HBeAb（+）血清中,HBV-LP阳性率（64.2%）明显高于HBV-DNA阳性率（P＜0.05）,而PreS1-Ag阳性率与HBV-DNA阳性率差异无统计学意义（P＞0.05）;65例HBeAg（－）HBeAb（－）血清中,HBV-LP阳性率（72.3%）和PreS1-Ag阳性率（67.7%）均明显高于HBV-DNA阳性率（Ps＜0.05）;（2）以185例HBeAg（－）乙肝患者的HBV-DNA表达量为参考标准,HBV-LP、PreS1-Ag的阳性预测值分别为72.6%、71.6%,阴性预测值分别为93.4%、84.1%;HBV-LP和PreS1-Ag联合检测,HBV-LP/PreS1-Ag双阳性中的HBV-DNA阳性率（66.0%）显著高于HBV-LP/PreS1-Ag双阴性（P＜0.05）。结论 血清HBV-LP和PreS1-Ag水平与HBV-DNA表达量有关,二者联合检测可灵敏地反映HBeAg（－）乙肝患者HBV的复制状态,预测HBV-DNA水平。     

An improved reverse dot hybridization for simple and rapid detection of adefovir dipivoxil-resistant hepatitis B virus

Early detection of adefovir dipivoxil-resistant mutants during long-term treatment of chronic hepatitis B virus (HBV) infection with this drug is of great clinical importance. We developed an improved reverse dot hybridization test for simple and rapid detection of the rtA181V/T and rtN236T mutations associated with adefovir dipivoxil resistance in chronic hepatitis B patients. Probes were designed for genotypes B, C, and D of this resistance characteristic; a total of 70 clinical samples were analyzed with this improved reverse dot hybridization assay. Its usefulness was validated by comparing with sequencing data. Discordant results were confirmed by subclone sequencing. This reverse dot hybridization assay was sufficiently sensitive to detect 10(3) copies/mL; it also detected adefovir dipivoxil-resistant mutant strains when they comprised more than 5% of a mixed virus population. This reverse dot hybridization array correctly identified adefovir dipivoxil-resistant mutants; it had high concordance (98.5%) with direct sequencing data. There was no clear relationship between the HBV genotype and the development of adefovir dipivoxil-resistant mutants. This reverse dot hybridization assay proved to be simple and rapid for detection of rtA181V/T and rtN236T mutations associated with resistance to adefovir dipivoxil.     

Pathological femoral fractures due to osteomalacia associated with adefovir dipivoxil treatment for hepatitis B: a case report

ABSTRACT: We present a case of a 62-year-old man who underwent total hip arthroplasty for treatment of pathologic femoral neck fracture associated with adefovir dipivoxil-induced osteomalacia. He had a 13-month history of bone pain involving his shoulders, hips, and knee. He received adefovir dipivoxil for treatment of lamivudine-resistant hepatitis B virus infection for 5 years before the occurrence of femoral neck fracture. Orthopedic surgeons should be aware of osteomalacia and pathological hip fracture caused by drug-induced renal dysfunction, which results in Fanconi's syndrome.Virtual slidesThe virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1600344696739249.     

Cellular immune response to hepatitis B virus-encoded antigens in acute and chronic hepatitis B virus infection

The proliferative response of PBMC to hepatitis B virus (HBV) envelope, core, and e Ag was analyzed prospectively in 21 patients with acute self-limited HBV infection and compared with the response of patients with chronic HBV infection and different levels of HBV replication (i.e., hepatitis e Ag (HBeAg)- or anti-HBe-positive) and liver damage (i.e., chronic active hepatitis or chronic asymptomatic carriers). Our results indicate that: 1) HBV-infected subjects who develop a self-limited acute hepatitis show a vigorous PBMC response to hepatitis B core Ag and HBeAg, as expression of T cell activation; 2) appearance of a detectable lymphocyte response to HBV nucleocapsid Ag is temporally associated with the clearance of HBV envelope Ag; 3) in patients with chronic HBV infection the level of T cell responsiveness to hepatitis B core Ag and to HBeAg is significantly lower than that observed during acute infection; 4) T cell sensitization to HBV envelope Ag in acute and chronic HBV infection is usually undetectable and when measurable is expressed transiently and at low levels. These results may reflect immune events of pathogenetic relevance with respect to evolution of disease and viral clearance.     

内江地区慢性乙型肝炎患者的基因型及拉米夫定联合阿德福韦酯抗病毒治疗效果

目的 探讨内江地区慢性乙型肝炎患者的基因型及拉米夫定联合阿德福韦酯抗病毒的治疗效果。方法 将201例慢性乙型肝炎患者进行HBV基因型的测定,其中120例HBeAg（+）慢性乙型肝炎患者随机分为三组：A组、B组和C组,每组40例。A组给予拉米夫定(LAM)治疗;B组给予恩替卡韦(ETV)治疗;C组给予LAM联合阿德福韦酯(ADV)治疗,比较治疗情况。结果 201例慢性乙型肝炎患者中B型119例（59.2%）,C型68例（33.8%）,B/C混合型10例（5.0%）,未知型4例（2.0%）,各分型之间的性别、年龄差异无统计学意义（P＞0.05）。C型感染者HBeAg阳性率为86.8%,显著高于B型的51.3%（P＜0.05）。HBeAg（+）的3组患者治疗12、24和48周时,B组和C组患者的ALT复常率、HBV DNA阴转率及48周时的HBeAg血清转换率均显著高于A组（P＜0.05）;C组患者与B组比较差异无统计学意义（P＞0.05）。治疗期间均未见不良反应发生。C组患者（包括B型19例、C型21例）中B型的HBV DNA阴转率及HBeAg血清转换率显著高于C型（P＜0.05）。结论 地处西南方的内江地区慢性乙型肝炎患者主要以B型为主,C型次之,B型和C型共占93.0%,其他型别仅占较少部分。ETV方案或LAM联合ADV方案治疗HBeAg（+）慢性乙肝疗效优于LAM治疗。初始LAM联合ADV治疗基因B型HBeAg（+）慢性乙型肝炎疗效优于C型。     

S-adenosyl-methionine and betaine improve early virological response in chronic hepatitis C patients with previous nonresponse

Background/Aims

Treatment of chronic hepatitis C (CHC) with pegylated interferon α (pegIFNα) and ribavirin results in a sustained response in approximately half of patients. Viral interference with IFNα signal transduction through the Jak-STAT pathway might be an important factor underlying treatment failure. S-adenosyl-L-methionine (SAMe) and betaine potentiate IFNα signaling in cultured cells that express hepatitis C virus (HCV) proteins, and enhance the inhibitory effect of IFNα on HCV replicons. We have performed a clinical study with the aim to evaluate efficacy and safety of the addition of SAMe and betaine to treatment of CHC with pegIFNα/ribavirin.

Methods

In this open-label pilot study, 29 patients with CHC who failed previous therapy with (peg)IFNα/ribavirin were treated with SAMe, betaine, pegIFNα2b and ribavirin. Treatment duration was 6 or 12 months, depending on genotype, and the protocol comprised a stopping rule at week 12 if early virological response (EVR) was not achieved. Virological and biochemical response and safety were assessed throughout the treatment.

Results

29 patients were enrolled and treated according to the study protocol. 79% of the patients were infected with genotype 1, 72% had advanced fibrosis, 76% had previously received pegIFNα/ribavirin, and only 14% achieved EVR to the previous treatment. When treated with the study medications, 17 patients (59%) showed an EVR, only 3 (10%) however achieved a sustained virological response (SVR). SAMe and betaine were found to be safe when used with pegIFNα/ribavirin.

Conclusion

The addition of SAMe and betaine to pegIFNα/ribavirin improves early virological response in CHC.

Trial Registration

ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT00310336","term_id":"NCT00310336"}}NCT00310336     

Transplacentally acquired maternal antibody against hepatitis B surface antigen in infants and its influence on the response to hepatitis B vaccine

Background

Passively acquired maternal antibodies in infants may inhibit active immune responses to vaccines. Whether maternal antibody against hepatitis B surface antigen (anti-HBs) in infants may influence the long-term immunogenicity of hepatitis B vaccine remains unknown.

Methodology/Principal Findings

Totally 338 pairs of mothers and children were enrolled. All infants were routinely vaccinated against hepatitis B based on 0-, 1- and 6-month schedule. We characterized the transplacental transfer of maternal anti-HBs, and compared anti-HBs response in children of mothers with or without anti-HBs. In a prospective observation, all 63 anti-HBs positive mothers transferred anti-HBs to their infants; 84.1% of the infants had higher anti-HBs concentrations than their mothers. One and half years after vaccination with three doses of hepatitis B vaccine, the positive rate and geometric mean concentration (GMC) of anti-HBs in 32 infants with maternal anti-HBs were comparable with those in 32 infants without maternal antibody (90.6% vs 87.5%, P = 0.688, and 74.5 vs 73.5 mIU/ml, P = 0.742, respectively). In a retrospective analysis, five and half years after vaccination with three doses vaccine, the positive rates of anti-HBs in 88 children of mothers with anti-HBs ≥1000 mIU/ml, 94 children of mothers with anti-HBs 10–999 mIU/ml, and 61 children of mothers with anti-HBs <10 mIU/ml were 72.7%, 69.2%, and 63.9% (P = 0.521), respectively; anti-HBs GMC in these three groups were 38.9, 43.9, and 31.7 mIU/ml (P = 0.726), respectively.

Conclusions/Significance

The data demonstrate that maternal anti-HBs in infants, even at high concentrations, does not inhibit the long-term immunogenicity of hepatitis B vaccine. Thus, current hepatitis B vaccination schedule for infants will be still effective in the future when most infants are positive for maternal anti-HBs due to the massive vaccination against hepatitis B.     

Association of polymorphisms in the interleukin-4 gene with response to hepatitis B vaccine and susceptibility to hepatitis B virus infection: A meta-analysis

Objective

The aim of this meta-analysis is to evaluate the associations between functional polymorphisms in the interleukin-4 (IL4) gene and individuals' responses to hepatitis B vaccine and their susceptibility to hepatitis B virus (HBV) infection.

Methods

A literature search on articles published before December 1st, 2012 was conducted in PubMed, Embase, Web of Science and China BioMedicine (CBM) databases. Crude odds ratios (ORs) with 95% confidence intervals (CIs) were calculated. Statistical analyses were performed using the STATA 12.0 software.

Results

Eight studies were eligible for inclusion in this meta-analysis, including five cross-sectional studies on individual's response to hepatitis B vaccine and three case–control studies on HBV infection risk. The meta-analysis results showed that the T allele of rs2243250, the T allele of rs2070874, and the C allele of rs2227284 in IL4 gene were associated with high responses to hepatitis B vaccine. Further subgroup analysis by ethnicity showed that there was a significant association between IL4 genetic polymorphisms and an individual's responses to hepatitis B vaccine among Asian populations, but similar association was not found among Caucasian populations. However, there was no evidence indicating a correlation between IL4 genetic polymorphism and susceptibility to HBV infection.

Conclusion

Our current meta-analysis suggests that rs2243250, rs2070874 and rs2227284 polymorphisms in IL4 gene may play an important role in determining the response to hepatitis B vaccine, especially among Asian populations. However, further studies are still needed to evaluate the associations between IL4 genetic polymorphisms and HBV infection risk.     

Development of a nasal vaccine for chronic hepatitis B infection that uses the ability of hepatitis B core antigen to stimulate a strong Th1 response against hepatitis B surface antigen

There are estimated to be 350 million chronic carriers of hepatitis B infection worldwide. Patients with chronic hepatitis B are at risk of liver cirrhosis with associated mortality because of hepatocellular carcinoma and other complications. An important goal, therefore, is the development of an effective therapeutic vaccine against chronic hepatitis B virus (HBV). A major barrier to the development of such a vaccine is the impaired immune response to HBV antigens observed in the T cells of affected patients. One strategy to overcome these barriers is to activate mucosal T cells through the use of nasal vaccination because this may overcome the systemic immune downregulation that results from HBV infection. In addition, it may be beneficial to present additional HBV epitopes beyond those contained in the traditional hepatitis B surface antigen (HbsAg) vaccine, for example, by using the hepatitis B core antigen (HBcAg). This is advantageous because HBcAg has a unique ability to act as a potent Th1 adjuvant to HbsAg, while also serving as an immunogenic target. In this study we describe the effect of coadministration of HBsAg and HBcAg as part of a strategy to develop a more potent and effective HBV therapeutic vaccine.     

Predictive value of early virological response to treatment with different interferon-based regimens plus ribavirin in patients with chronic hepatitis C

Early virological response (EVR) to different interferon-based regimens plus ribavirin and its ability to predict the outcome of therapy in patients with chronic hepatitis C were investigated. The study design was as follows: 64 naive patients were considered, 32/64 received pegylated interferon alpha-2b (Peg-IFN-alpha2b) plus ribavirin and the remaining 32 received leucocyte interferon alpha (IFN-alpha) plus ribavirin. At week 4 of treatment, EVR was present in 68.7% and 37.5% of patients treated with Peg-IFN-alpha2b plus ribavirin, and with leucocyte interferon alpha (IFN-alpha) plus ribavirin, respectively (p = 0.024). At week 12, the cumulative EVR rates did not differ between the two groups (71.9% vs 56.2%, p >0.05) because a higher proportion of patients achieved EVR for the first time after more than 4 weeks of therapy in the standard IFN-alpha group. Sustained virological response (SVR) rates, however, resulted significantly higher in the Peg-IFN-alpha2b group (65.6% vs 37.5%; p = 0.045) since a higher proportion of patients who received standard IFN-alpha relapsed during the follow-up. In the standard IFN-alpha group, HCV genotype 1 (p = 0.035), high baseline viral load (p = 0.035) and the presence of bridging fibrosis/cirrhosis (p = 0.011) were closely associated with significantly lower SVR rates. In the Peg-IFN-alpha2b group, only bridging fibrosis/cirrhosis (p = 0.02) negatively influenced the outcome of treatment. Overall, 33/41 (80.5%) patients with EVR at week 12 were sustained responders, yielding a positive predictive value (PPV) of 0.80. However, when SVR was related to the time taken to reach EVR, 32/34 (94.1%) patients with EVR at week 4 of therapy (PPV = 0.94) versus 1/7 (14.3%) patients who had EVR after more than 4 weeks of therapy (PPV = 0.14) resulted sustained responders (p = 0.000057). In conclusion, EVR at week 4 of treatment is strongly associated with the likelihood of achieving SVR, regardless of the therapeutic regimen. However, when compared with standard IFN-alpha plus ribavirin, treatment with Peg-IFN-alpha2b plus ribavirin significantly increases the probability of viral clearance within the first 4 weeks of treatment. Finally, patients who do not clear the virus within the first 12 weeks of treatment have no chance of achieving SVR, justifying discontinuation of therapy in these patients.     

Low immune response to hepatitis B vaccine among children in Dakar, Senegal

HBV vaccine was introduced into the Expanded Programme on Immunization (EPI) in Senegal and Cameroon in 2005. We conducted a cross-sectional study in both countries to assess the HBV immune protection among children. All consecutive children under 4 years old, hospitalized for any reason between May 2009 and May 2010, with an immunisation card and a complete HBV vaccination, were tested for anti-HBs and anti-HBc. A total of 242 anti-HBc-negative children (128 in Cameroon and 114 in Senegal) were considered in the analysis. The prevalence of children with anti-HBs ≥ 10 IU/L was higher in Cameroon with 92% (95% CI: 87%-97%) compared to Senegal with 58% (95% CI: 49%-67%), (p<0.001). The response to vaccination in Senegal was lower in 2006-2007 (43%) than in 2008-2009 (65%), (p = 0.028). Our results, although not based on a representative sample of Senegalese or Cameroonian child populations, reveal a significant problem in vaccine response in Senegal. This response problem extends well beyond hepatitis B: the same children who have not developed an immune response to the HBV vaccine are also at risk for diphtheria, tetanus, pertussis (DTwP) and Haemophilus influenzae type b (Hib). Field biological monitoring should be carried out regularly in resource-poor countries to check quality of the vaccine administered.     

Impaired humoral immune response to hepatitis B vaccine in patients on maintenance hemodialysis

Hepatitis B virus (HBV) infection is a worldwide health problem. We aimed in this study to investigate the humoral immune response derived to HBV vaccine following completing the vaccine series in Madinah. Two hundred and two Saudi hemodialysis (HD) patients were included in this cross-sectional study. Mean concentration of Hepatitis B surface antibody (anti-HBs) was significantly higher among patients who received the vaccination twice compared to patients who received the vaccination only after starting hemodialysis (252 ± 489 mIU/mL vs. 144 ± 327 mIU/mL, respectively, p = 0.008). Almost half of the study sample were non-protected and showed anti-HBs concentration < 10 mlU/mL. In contrast, 20.3% (n = 41) were identified as poor responders (10–100 mlU/mL) and only 28.2% (n = 57) were identified as good responders (10–100 mlU/mL). However, the latter two groups were accounted as protected (48.5%, n = 98). Patients sex was associated with anti-HBs concentration (non-responders; poor responders; good responders), where significantly higher proportion of good responders were females compared to males (p = 0.007). In conclusion, HBV vaccine is efficient to elicit humoral immune response in hemodialysis patients.     

Role of immunoproteasome catalytic subunits in the immune response to hepatitis B virus

Inhibition of hepatitis B virus (HBV) replication and viral clearance from an infected host requires both the innate and adaptive immune responses. Expression of interferon (IFN)-inducible proteasome catalytic and regulatory subunits correlates with the IFN-alpha/beta- and IFN-gamma-mediated noncytopathic inhibition of HBV in transgenic mice and hepatocytes, as well as with clearance of the virus in acutely infected chimpanzees. The immunoproteasome catalytic subunits LMP2 and LMP7 alter proteasome specificity and influence the pool of peptides available for presentation by major histocompatibility complex class I molecules. We found that these subunits influenced both the magnitude and specificity of the CD8 T-cell response to the HBV polymerase and envelope proteins in immunized HLA-A2-transgenic mice. We also examined the role of LMP2 and LMP7 in the IFN-alpha/beta- and IFN-gamma-mediated inhibition of virus replication using HBV transgenic mice and found that they do not play a direct role in this process. These results demonstrate the ability of the IFN-induced proteasome catalytic subunits to shape the HBV-specific CD8 T-cell response and thus potentially influence the progression of infection to acute or chronic disease. In addition, these studies identify a potential key role for IFN in regulating the adaptive immune response to HBV through alterations in viral antigen processing.     

Discrepancy in virological and biochemistry response of patients with chronic hepatitis HCV positive on treatment with PEG-IFN plus ribavirin

Combination therapy of PEG-IFN alpha-2a o alpha-2b plus ribavirin represents a further improvement in treatment of chronic hepatitis HCV+ with a sustained virological response (SVR) either in monotherapy (25-39%) either in association with ribavirin (59-56%). SVR is highly predictable: 75% of all patients who achieve viral clearance at week 12 (EVR), if they had an adherence > 80% of planned therapy, they become sustained viral responders. In spite of virological response, 16-34% of patients on PEG-IFN monotherapy have high value of ALT, and this make them to reduce adherence. 62 patients whith chronic hepatitis HCV+ and no corrhosis, have been treated for 48 weeks with PEG-IFN and ribavirin to evaluate discrepancy incidence between virological (HCVRNA < 200UI) and biochemical (normal value of ALT) response of patients treated with PEG-IFN plus ribavirin and to verify the impact that stuch discrepancy can produce on SVR of treated patients. Our preliminary data confirm that PEG-IFN bring a superior virological response than biochemical one, either on naive patients either on experienced ones even with ribavirin in association. It will be useful to verify if this discepancy cause a superior SVR as already reported by several studies. Even the follow-up of our 5 discordant patients confirm this trend.     

硫普罗宁治疗慢性乙型肝炎的临床疗效对照研究

目的：评价硫普罗宁治疗慢性乙型肝炎的效果及安全性。方法：慢性乙型肝炎80例,随机分为两组。治疗组40例,以硫普罗宁0.2g加入5%葡萄糖溶液250ml中静滴,1次/日,连续治疗4周,对照组40例,以甘草酸二铵150mg加入5%葡萄糖溶液250ml中静滴,1次/日,连续治疗4周;同时分别监测肝功能指标：丙氨酸氨基转移酶（ALT）,天冬氨酸转移酶（AST）、总蛋白（TP）、总胆红素（TBi）、肝纤维化指标（HA、PCⅢ）及凝血酶原活动度（PTA）等,并进行比较分析。结果：硫普罗宁治疗组较对照组在肝功能复常方面显示良好的治疗效应,可使转氨酶及HA明显下降,两组显效率分别为50%（20/40）和25%（10/40）,总有效率为85%（34/40）和47.5%（19/40）,两组比较差异有显著性（p〈0.01）。结论：短期临床研究提示硫普罗宁治疗慢性乙型肝炎,可明显改善肝脏功能,有抗纤维化作用,且副作用少,疗效显著。