Role of Immune Cells in Hepatitis B Virus and Associated Sequelae.

Natural history of chronic hepatitis B in Euro-Mediterranean and African Countries

Objective The study aims to investigate the associationbetweencholesterol 7α-hydroxylase (CYP7A1) gene polymorphism and different clinical outcomes after Hepatitis B virus (HBV) infection in Fujian Han population and lay a foundation for understanding the mechanisms of genesis and development of HBV-related diseases. Methods Case-control study was conducted.586 patients of HBV persistent infection without antiviral therapy and 225 HBV rehabilitation patients(35-55 years old) were collected from May 2015 to June 2016 in the Liverish Center of First Clinical College of Fujian Medical University.The group of HBV persistent infection without antiviral therapy included 246 patients with chronic hepatitis B, 177 patients with hepatitis B-related cirrhosis, and 163 patients with hepatitis B-related liver cancer.The rs3824260, rs4738687and rs8192871 loci of CYP7A1 gene were detected by improved multiple ligase detection reaction (iMLDR). Logistic regression analysis and chi-square test were used to analyze the genotyping results. Results Three SNPs (single nucleotide polymorphisms) of CYP7A1 gene were selected and compared between HBV persistent infection group and HBV rehabilitation group and between chronic hepatitis B subgroup, liver cirrhosis subgroup and liver cancer subgroup. After adjustment for factors including age andgender, there was no significant difference in the distribution of rs3824260 genotype among the groups(χ2=1.565, P=0.459), however, the frequency of allele C in HBV rehabilitation group was significantly higher than in HBV persistent in fectiongroup for men(χ2=4.365, P=0.037), whereas the frequency of rs3824260 CC and CT was more likely to be observed in liver cancer group than in non-liver cancer group (chronic hepatitis B subgroup and liver cirrhosis subgroup) for women (χ2=5.768, P=0.012; χ2=10.130, P=0.001). The frequency of rs4738687 GG genotype was more likely to be observed in non-liver cancer group than in liver cancer group (χ2=4.403, P=0.041; χ2=6.940, P=0.009). The results of gender stratification showed that there were significant differences in the distribution of rs4738687 among the HBV persistent infection groups for men(χ2=10.697, P=0.030), however, there was no significant difference in the distribution of rs4738687 among the HBV persistent infection groups for women(χ2=4.627, P=0.329), and there was no significant difference in the distribution of genotype frequency and allele frequency among all groups(χ2=0.489, P=0.792). There was no significant difference after sex stratification either(χ2=1.282, P=0.526; χ2=1.565, P=0.465). Conclusions These findings suggested that CYP7A1 gene polymorphism was related todifferent clinical outcomes in Fujian Han population. The rs3824260 mutation had a certain gender preference and the mutation allele was detected in a higher proportion in male patients. Male HBV patients with rs3824260 C allele had more chance of switching to rehabilitation. The rs4738687 was likely to be related to the occurrence of liver cancer in Fujian Han population, and GG genotype may delay the occurrence and development of liver cancer especially in the male group. The rs8192871 was not found to be related to the different clinical outcomes of HBV infection.(Chin J Lab Med, 2018, 41: 155-164) Key words: Hepatitis B virus; Polymorphism single nucleotide; Cholesterol 7-alpha-Hydroxylase

Objective To study the relationship between hepatitis B virus genotypes and hepatitis B virus large envelope protein. Methods A total of 138 surveyed children with chronic active HBV infection were enrolled real-time fluorescence PCR method was employed to quantify serum HBV DNA levels and classify the HBV genotypes. Hepatitis B virus large envelope protein was detected by using enzyme linked immuno sorbent assay (ELISA). Results The absorbance of hepatitis B virus large envelope protein was positively correlated with hepatitis B virus DNA levels (r=0.85, P＜0.05); Serum ALT levers, the absorbance of hepatitis B virus large envelope protein and hepatitis B virus DNA levels (P＞0.05,P＞0.05,P＞0.05), were no significant difference between hepatitis B virus genotype B and hepatitis B virus genotype C. Conclusion There was a close correlation between the absorbance of hepatitis B virus large envelope protein and HBV DNA levels. Hepatitis B virus large envelope protein was a reliable serological marker that can reflect the replication of hepatitis B virus and there was no relationship between hepatitis B virus genotypes and Hepatitis B virus large envelope protein. Key words: Hepatitis B, Chronic; Genotype; Viral envelope proteins

To comprehensively measure the burden of hepatitis B, liver cirrhosis and liver cancer in Shandong province, using disability-adjusted life years (DALYs) to estimate the disease burden attribute to hepatitis B virus (HBV) infection. Based on the mortality data of hepatitis B, liver cirrhosis and liver cancer derived from the third National Sampling Retrospective Survey for Causes of Death during 2004 and 2005, the incidence data of hepatitis B and the prevalence and the disability weights of liver cancer gained from the Shandong Cancer Prevalence Sampling Survey in 2007, we calculated the years of life lost (YLLs), years lived with disability (YLDs) and DALYs of three diseases following the procedures developed for the global burden of disease (GBD) study to ensure the comparability. The total burden for hepatitis B, liver cirrhosis and liver cancer were 211,616 (39,377 YLLs and 172,239 YLDs), 16,783 (13,497 YLLs and 3286 YLDs) and 247,795 (240,236 YLLs and 7559 YLDs) DALYs in 2005 respectively, and men were 2.19, 2.36 and 3.16 times as that for women, respectively in Shandong province. The burden for hepatitis B was mainly because of disability (81.39%). However, most burden on liver cirrhosis and liver cancer were due to premature death (80.42% and 96.95%). The burden of each patient related to hepatitis B, liver cirrhosis and liver cancer were 4.8, 13.73 and 11.11 respectively. Hepatitis B, liver cirrhosis and liver cancer caused considerable burden to the people living in Shandong province, indicating that the control of hepatitis B virus infection would bring huge potential benefits.

Liver cancer is the second leading cause of cancer death worldwide, responsible for an estimated 746,000 deaths and 782,000 new cases in 2012 [1]. The countries in the Western Pacific region accounts for 64% and China alone accounts for 51% of the new liver cancer cases and deaths each year. Approximately 80% of hepatocellular carcinoma (HCC), the most common type of liver cancer, is associated with viral hepatitis [1]. In countries with high prevalence of hepatitis B virus (HBV) infection, such as China, up to 80% of HCC is associated with hepatitis B [2]. Liver cancer carries a poor prognosis with a global mortality to incidence ratio of 0.95 [1]. In the USA, the 1-year survival rate remains less than 50% [3]. Asians and Pacific Islanders have the highest incidence of HCC among the different racial/ethnic groups. While every individual with chronic hepatitis B infection (CHB) or HBV carrier is at risk for developing HCC, disease progression leading to cirrhosis is associated with the greatest risk [2]. In an effort to improve survival through early detection of HCC, the American Association for the Study of Liver Diseases recommended HCC screening with abdominal ultrasound (US) at 6–12 month intervals of HBV carriers considered at increased risk for HCC, including those with cirrhosis, family history of liver cancer and Asian male HBV carriers above the age of 40 [4,5]. However, it remains controversial whether population-wide HCC screening of CHB patients results in a reduction in HCC mortality particularly in resource-constraint regions of the world that have the highest burden of HCC. The only two randomized trials available to date are both from China [6,7]. A population-based study in the city of Shanghai, conducted by the Liver Cancer Institute of Fudan University, reported 1-year HCC survival rates improved from 31.2% in the control group to 65.9% in the screened group among 18,816 CHB patients aged 35–59 years with or without cirrhosis, who were randomized into either a group using US and AFP screening every 6 months or a control group [6]. The second study from a rural setting in Qidong, using AFP screening every 6 months without US, did not show any improvement in survival [7]. According to the National Cancer Institute [8], screening for HCC

“Functional cure” is being pursued as the ultimate endpoint of antiviral treatment in chronic hepatitis B (CHB), which is characterized by loss of HBsAg whether or not anti-HBs antibodies are present. “Functional cure” can be achieved in <10% of CHB patients with currently available therapeutic agents. The dysfunction of specific immune responses to hepatitis B virus (HBV) is considered the major cause of persistent HBV infection. Thus, modulating the host immune system to strengthen specific cellular immune reactions might help eliminate HBV. Strategies are needed to restore/enhance innate immunity and induce HBV-specific adaptive immune responses in a coordinated way. Immune and resident cells express pattern recognition receptors like TLRs and RIG I/MDA5, which play important roles in the induction of innate immunity through sensing of pathogen-associated molecular patterns (PAMPs) and bridging to adaptive immunity for pathogen-specific immune control. TLR/RIG I agonists activate innate immune responses and suppress HBV replication in vitro and in vivo, and are being investigated in clinical trials. On the other hand, HBV-specific immune responses could be induced by therapeutic vaccines, including protein (HBsAg/preS and HBcAg), DNA, and viral vector-based vaccines. More than 50 clinical trials have been performed to assess therapeutic vaccines in CHB treatment, some of which display potential effects. Most recently, using genetic editing technology to generate CAR-T or TCR-T, HBV-specific T cells have been produced to efficiently clear HBV. This review summarizes the progress in basic and clinical research investigating immunomodulatory strategies for curing chronic HBV infection, and critically discusses the rather disappointing results of current clinical trials and future strategies.

Objective To investigate the clinical features of hepatitis B virus (HBV) reactivation after precise radiotherapy in patients with primary liver cancer (PLC) and analyze the risk factors for HBV reactivation.Methods A retrospective analysis was performed on the clinical data of 69 hepatitis B surface antigen (HBsAg)-positive patients with PLC,some of whom had HBV reactivation after precise radiotherapy.Before radiotherapy,all patients underwent baseline examinations,including blood routine,liver function test,renal function test,and quantifications of serum alpha-fetoprotein (AFP),serum HBV markers,and serum HBV DNA.During radiotherapy and within 12 weeks after radiotherapy,blood routine was performed biweekly,and liver function test,renal function test,and quantifications of serum AFP,serum HBV markers,and serum HBV DNA were performed once every four weeks.Logistic regression analysis was used to evaluate the association of the indices with HBV reactivation.Results Of the 69 patients,12 (17％) had radiation-induced liver disease,17 (25 ％) had HBV reactivation,and 15 (22％)developed hepatitis due to HBV reactivation.The logistic regression analysis showed that baseline serum HBV DNA level was the risk factor for HBV reactivation after precise radiotherapy.Conclusions HBV reactivation may occur after precise radiotherapy in patients with PLC,and baseline serum HBV DNA level is the independent risk factor for HBV reactivation.The patients who develop hepatitis due to HBV reactivation have poor prognosis even if they receive antiviral therapy in time. Key words: Liver neoplasm/three-dimensional conformal radiotherapy; Radiation-induced liver disease; Hepatitis B virus

Pre-S Deletion and Complex Mutations of Hepatitis B Virus Related to Advanced Liver Disease in HBeAg-Negative Patients

Objective To investigate the expression and clinical significance of ribonucleotide reductase small subunit M2 (RRM2) in chronic hepatitis B virus (HBV) infection and related liver diseases. Methods A total of 428 patients with chronic HBV infection and liver disease were enrolled from Songyang County People’s Hospital from October 2017 to September 2019. There were 166 cases of chronic hepatitis B (CHB), 53 cases of HBV-related cirrhosis, 28 cases of non-HBV-related cirrhosis, 57 cases of HBV-related liver cancer, 33 cases of non-HBV-related liver cancer, and 91 cases of non-viral hepatitis. In addition, 36 healthy subjects were selected as the control group. Among 166 cases of CHB, there were 87 patients with high viral load group (HBV DNA ≥4.0 lg IU/mL) and 79 patients with low viral load group (HBV DNA <4.0 lg IU/mL); while in 87 high viral load patients, 56 had high alanine transaminase (ALT) (≥40 U/L) and 31 had normal ALT (<40 U/L). The expression level of serum RRM2 protein in patients was detected by enzyme-linked immunosorbent assay (ELISA), and the relationship of RRM2 expression with HBV DNA and liver function was analyzed. SPSS 23.0 and PRISM 8.0 statistical software were used to analyze data. Correlation analysis was performed using Spearman analysis. Results The serum ALT and RRM2 levels in patients with high viral load CHB were higher than those in low viral load group (Z=-6.68, t=6.80, P 0.05). The level of ALT in the non-viral hepatitis group was higher than that in the healthy control group (Z=-8.43, P 0.05). Correlation analysis showed that serum RRM2 level was positively correlated with HBV DNA load (r=0.51, P 0.05) in patients with chronic HBV infection and related liver diseases. Conclusions Serum RRM2 level is positively correlated with HBV DNA load and has no significant correlation with ALT. RRM2 might be used as a target for the development of new hepatitis B drugs. Key words: Hepatitis B, chronic; HBV DNA; Ribonucleotide reductase; Alanine transaminase

Scaling up hepatitis B vaccination with the support of GAVI in China : lessons learned for introduction of new vaccines and for the future of hepatitis B control

To investigate the relationship of alcohol intake and hepatocellular carcinoma among patients with hepatitis B virus infection. A total of 553 patients with HCC and 160 control subjects affected with hepatitis B virus were recruited. Serum virology, serum biochemistry, as well as demographic information were studied. Finally, risk factors were selected by stepwise Logistic regression analyse. Odds ratios (ORs) were estimated for each risk factor. According to alcohol intake, HCC patients were divided into three groups,then to observe the differences between them. Elevated AST, GGT, ALP and AFP levels were seen more frequently in the HCC case groups compared to control group (P < 0.05). Multivariate analysis revealed that heavy alcohol use, smoking, positive family history of liver cancer is associated with HCC development among patients with hepatitis B virus infection. Significantly increased risk was found among patients for heavy alcohol use [A = 2.66 (2.01-3.50)] and for smoking [A = 2.51 (1.66-3.80)] and for positive family history of liver cancer [A = 1.64 (1.04-2.59)]. Compared to patients who did not have alcohol use, elevated GGT and ALP were seen more frequently in patients who had alcohol use either mild or heavy (P < 0.05). Heavy alcohol use, smoking, positive family history of liver cancer is positive correlation with HCC development among patients with hepatitis B virus infection in China. In patients with hepatitis B virus infection who also has history of heavy alcohol, the most risk factor of HCC is hepatitis B virus infection, not alcohol.

Objective To evaluate the efficacy of tebivudine in the treatment for maternal-infant blockade of pregnant women with both serum hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) positive and high load of hepatitis B virus (HBV) infection in late pregnancy. Methods From July 1st, 2007 to June 30th, 2017, pregnant women with chronic HBV infection in late pregnancy (28 weeks of pregnancy) and babies delivered in the outpatient Department of Gynaecology and Obstetrics, Beijing Ditan Hospital, Capital Medical University were selected. Total of 250 pregnant women with both HBsAg and hepatitis B e antigen (HBeAg) positive, normal liver function, HBV DNA ≥ 1 × 106 copies/ml and with complete follow-up data were collected. The pregnant women were fully informed of the advantages and disadvantages of the maternal-infant blocking of the anti-viral medication during pregnancy. According to the voluntary principle, the pregnant women were divided into tebivudine group (150 cases) and control group (100 cases) according to whether treated with tebivudine. The pregnant women in tebivudine group received oral tebivudine 600 mg/d, from 28 weeks of pregnancy to 42 days after delivery. The control group did not use antiviral drugs. The serum HBV DNA load were detected in both groups at 28 weeks and at the time of delivery. The adverse reactions of pregnant women in tebivudine group after taking medicine were observed. The infants of both groups were immunized with active and passive immunization after birth; hepatitis B vaccine was injected intramuscularly within 6 hours, 1 month and 6 months, respectively, while intramuscular injection of hepatitis B immunoglobulin for 200 IU. The positive rate of HBsAg and the load of HBV DNA in 6 hours after the bith and at 7-month of the baby were detected (venous blood was drawn before passive immunization). Results Compared with the control group, the average HBV DNA load was significantly lower in tebivudine group after treatment and before delivery, with significant difference (t = 31.07, P < 0.001). The positive rate of serum HBV DNA in tebivudine group was significantly higher than that of control group within 6 hours of the babies birth, with significant difference (0.00% vs. 17.00%; χ2 = 27.36, P < 0.001). At the age of 7 months, the HBV DNA load of the infants in tebivudine group were all lower than the limit of detection, and the positive rate of HBV DNA in the control group was 10.00%. Fisher accurate test showed that the intrauterine infection rate of control group was significantly higher than that of tebivudine group (12.00% vs. 0.00%), with significant difference (P < 0.001). No serious adverse reactions were found during the follow-up period of tebivudine use, and no birth defects were found. Conclusions With the increase of serum HBV DNA load in pregnant women before delivery, the risk of HBV intrauterine infection increases. Normal liver function, both HBsAg and HBeAg positive, high HBV DNA load with tebivudine antiviral therapy in late of pregnancy could significantly reduce the serum HBV DNA load of pregnant women and block HBV intrauterine infection of effectively. It is safe for both mothers and infants to use tebivudine in late pregnancy. Key words: Telbivudine; Hepatitis B immunoglobulin; Hepatitis virus B; Hepatitis B surface antigen; Hepatitis B e antigen; Maternal-infant blockade; Intrauterine infection

IL-18 is a new type of IFN-γ inducible factor. It can not only trigger NK cells and Th1 cells to produce IFN-γ, but also promotes the proliferation of Th1 cells and NK cells and the activity of cytotoxinic function. As the impo rtant factors of regulating Th1 and Th2's cytokines, IL-18, IL-18 receptor (IL-18R) and IL-18 binding protein (IL-18BP) have currently been proved by experiments that they have strong relations with the pathogenesis of chronic hepatitis B. So they play significant roles in regulating the immune net of HBV infected patients. In the article, the researeb of IL-18 character and its signal transduction, the antiviral mechenisms and the association with HBV infection are reviwed. Key words: Interleukin-18; Hepatitis B, chronic; Th1 cells

Hepatitis B still becomes a big health problem in many countries including Indonesia. Transmission of the disease by the hepatitis B virus easily occurs. The virus can be identified in most of the body fluids such as saliva, semen, breast milk and serous cavity fluid. Every person has the risk of contracting Hepatitis B. The people do not realize that they are infected by the Hepatitis B virus, because their health is still in good condition and can transmit it to others. And for the patients, their condition can be worse because it can develop into chronic infection that can cause to cirrhosis and liver cancer. The purpose of the examination of the HBsAg for the people in Rt.06 of Sukajaya is to know how their health status concerning Hepatitis B infection. The method of this activity is carried out in a survey to determine the number of people as well as of their knowledge about Hepatitis B, and then will be held the examination of the HBsAg.The result of the examination of 44 people are known to all negative, but a survey of 185 people as many as 174 people are less aware of the danger of Hepatitis B virus. Therefore, it urges to give valuable information about the disease, and giving vaccination for people who do not have immunity power to exist Hepatitis B virus is highly recommended.

Objective To explore the changes of main indexes of liver function of peripheral blood, T cell subsets and the viral load in hepatitis B virus infection patients in different course and its clinical significance. Method 170 cases of hepatitis B virus infection patients were divided into acute hepatitis B group (18 cases), chronic hepatitis B group (73 cases), liver cirrhosis group (41 cases) , hepatic carcinoma group (38 cases), and normal control group of 40 cases according to the course. The related indexes of liver function and T cell subsets were detected by the automatic biochemical analyzer and flow cytometry, and the content of HBV-DNA was detected by real-time fluorescent quantitative PCR method. Results Alanine transaminase (ALT) and aspartate transaminase (AST) increased in four experimental groups compared to normal controls, with significant difference (P 0.05). The increase of alkaline phosphatase(ALP) was in liver cancer patients, the levels of ALP in chronic hepatitis B was low. Groups of total protein (TP) were within the normal reference range.CD3+ T cells in peripheral blood of the four experimental groups were lower than in the normal controls, the patients with hepatic carcinoma and cirrhosis was obviously lower. The proportions of CD4+ T cells rose in acute hepatitis group, but the percentage of CD8+ T cells is the lowest in acute hepatitis B patients.The most obvious rise of HBV DNA virus load in peripheral blood was in chronic hepatitis B group, but the HBV-DNA level is relatively low in acute hepatitis, liver cirrhosis and liver cancer patients, the lowest was in acute hepatitis B, and the differences were all statistically significant(P<0.05)compared with other groups. Conclusion The liver function, the expression of T cell subsets and HBV-DNA content had significant difference in hepatitis B virus infection patients with different course, the combined detection of these projects can provide guidance for the clinical assessment of the disease and course, and provide reliable basis for judging the prognosis of the disease and treatment. Key words: Hepatitis B virus; Liver function; HBV-DNA; T lymphocyte subsets