Correlation between HBsAg level and HBV DNA titer during chronic hepatitis B treatment

AIM: Viral load is widely used as an indicator for the diagnosis and monitoring the treatment efficacy of chronic hepatitis B (CHB). Previous studies suggested that the quantity of hepatitis B surface antigen (HBsAg) in serum could be a surrogate marker of serum hepatitis B virus (HBV) DNA level. In this study, we aimed to investigate whether HBsAg level correlates with HBV DNA titer during CHB treatment. METHODS: Sera were collected from 47 CHB patients ［35 male, 12 female, mean age: (35±8) years］ treated for 48 weeks with a monotherapy (pegylated interferon alpha-2a, 18 patients; lamivudine, 15 patients) or a combination therapy (pegylated interferon alpha-2a and lamivudine, 14 patients). Serum samples were obtained at week 0 (just before the treatment), 4, 8, 24, 48 and week 72 (24 weeks after the treatment). HBV DNA was measured with real-time polymerase chain reaction (PCR). HBsAg was quantified with an automated chemiluminescent microparticle immunoassay. RESULTS: The titer of HBsAg correlated with the HBV DNA level in the 18 patients with monotherapy of pegylated interferon alpha-2a and the 14 patients with combination therapy (pegylated interferon alpha-2a and lamivudine). The significant correlation (canonical correlation=0.83) was found. However, no correlation in 15 patients with the monotherapy of lamivudine was observed. CONCLUSION: HBsAg titer correlates with HBV DNA level in CHB patients during the treatment with interferon or interferon and lamivudine, which can be a surrogate marker for monitoring the treatment efficacy.     

Effects of ethanol on HBV gene mutations in patients with chronic hepatitis B by gene chip

AIM: To study ethanol influence on gene mutations of HBV DNA and to offer testimony for clinical diagnosis and treatment of chronic hepatitis B. METHODS: 85 patients with chronic hepatitis B were divided into alcoholic group and non-alcoholic group. Gene chip technique was used to detect gene mutations located in Pre-C nt G1896A and nt A1814C, basal core promoter (BCP) nt A1762T and nt G1764A, P nt C528A and nt T552C. RESULTS: The mutation frequency on BCP nt A1762T and nt G1764A in alcoholic group was significantly higher than that in non-alcoholic group (P<0.05). No difference of mutation frequency on pre-c nt G1896A nt A1814C and P nt C528A nt T552C between alcoholic and non-alcoholic group was observed (P>0.05). CONCLUSION: Ethanol stimulates HBV gene mutations on BCP nt A1762T and nt G1764A, enhances HBV DNA replication and gene expression, deteriorates the state of the illness.     

Effect of hepatitis virus B protein on the functions of PBMCs isolated from patients with chronic HBV infection

AIM: To study the effect of hepatitis virus B proteins on peripheral blood mononuclear cells (PBMCs) from patients among various types of chronic hepatitis B virus (HBV) infection.METHODS: 80 patients of various types of chronic HBV infection were observed, including 40 HBeAg positive with abnormal alanine aminotransferase (ALT) (A group), 20 HBeAg positive with persistent normal ALT(B group), 20 HBeAg and HBV-DNA negative with persistent normal ALT level(C group). IL-10, IFN-γ in CD8+CD28+T cells, after stimulation with PHA, HBeAg and HBcAg for 48 h, were inspected respectively in PBMCs.RESULTS: IFN-γ was significantly lower in HBeAg positive patients. IL-10 was significantly higher in HBeAg positive with normal ALT. CD8+CD28+T were significantly lower than others. CONCLUSION: In HBeAg positive group, secretion of cytotoxic T lymphocyte (CTL) and Th1 type cellular immunologic reaction is decreased, Th2 type cellular immunologic reaction is enhanced.     

Role of hepatitis B virus in intrahepatic TGF-β1/Smads signaling pathway

AIM:To explore the effects of hepatitis B virus (HBV) on intrahepatic expression of transforming growth factor β1（TGF-β1） and Smads. METHODS:The expression of intrahepatic TGF-β1, HBsAg and HBcAg in control group and chronic hepatitis B (CHB) group was detected by immunohistochemical method.The serum HBV DNA content was determined by real-time PCR. The role of HBV in the expression of TGF-β1, Smad3 and Smad7 in human hepatic stellate cell line LX-2 in vitro was observed by cell culture and Western blotting. RESULTS:The average score of intrahepatic TGF-β1 expression in CHB group was higher than that in control group. With the increase in serum HBV DNA content, intrahepatic TGF-β1 expression was also enhanced. In the HBcAg positive hepatic tissue, there was higher TGF-β1 expression than that in the liver tissue of HBcAg negative. Compared with control group and HBV+anti-TGF-β1 group, HBV caused increased expression of TGF-β1 and Smad3 in HBV group in vitro. No difference of Smad7 protein among control group, HBV group and HBV+anti-TGF-β1 group was observed. CONCLUSION: The expression of intrahepatic TGF-β1 is related to serum HBV DNA and hepatocellular HBcAg in the patients with CHB. HBV-induced liver fibrosis mainly relies on positive regulatory mechanisms of Smad3,and the negative regulation by Smad7 almost does not function.     

Correlation between clinical stages of HBV chronic infection and co-expression of CD244 and PD-1 in HBV-specific CTLs

AIM: To investigate the co-expression of cluster of differentiation 244 (CD244) and programmed death 1 protein (PD-1) on hepatitis B virus (HBV) antigen-specific cytotoxic T-lymphocytes (CTLs) in chronic hepatitis B (CHB) patients and its correlation with the clinical stages of HBV chronic infection. METHODS: Eighty-one CHB subjects with human leukocyte antigen-A2 (HLA-A2) positive, including 20 cases of acute-on-chronic liver failure (ACLF), 20 cases of severe chronic hepatitis B (s-CHB), 34 cases of mild and moderate chronic hepatitis B (m-CHB) and 7 cases of immune tolerant stage (IT) of chronic hepatitis B as well as 14 healthy controls (HC), were enrolled in this study. The co-expression of CD244 and PD-1 was analyzed in virus-specific CD8⁺ T-cells derived from peripheral blood using major histocompatibility complex class I (MHC-I) pentamers targeting immunodominant epitopes of HBV core antigen (18-27). RESULTS: In the patients with chronic HBV infection, virus-specific CD8+T-cells with co-expression of CD244 and PD-1 were at increased levels than those in total CD8+ T-cells (67.48%±17.16% vs 14.01%±7.97%, P<0.01) in the peripheral blood. Among different clinical stages, increased level of CD244 expression coincided with increased expression of PD-1 in m-CHB compared with IT of CHB (73.08%±8.63% vs 53.11%±18.05%, P<0.05), which was followed by decreased co-expression level in ACLF (63.11%±13.87% vs 72.05%±16.86%, P<0.05) and restoration of the ability to secrete IFN-γ (30.95%±20.29% vs 13.63%±10.46%, P<0.05). CONCLUSION: CD244 and PD-1 are highly co-expressed in HBV-specific CTLs in the patients with s-CHB and m-CHB, and are decreased in ACLF following the restoration of IFN-γ secretion. The severity of CHB may be correlated with CD244 and PD-1 co-expression in HBV-specific CTLs.     

Inhibitory effect of HMGN2 protein on human hepatitis B viral HBeAg and HBsAg expression and DNA replication in HBV-transformed HepG2. 2.15 cell line

AIM: We previously demonstrated that HMGN2 is an antibacterial effector molecule in human LAK cells. This study was to examine the antiviral activity of HMGN2 against human hepatitis virus.METHODS: The purification and identify of HMGN2 proteins including preparative acid-urea polyacymide gel electrophoresis elution, reverse-phase high-performance liquid chromatography, mass spectrum, Western blotting and antimicrobial assay were conducted. The cellular toxicity of HMGN2 to the HepG2.2.15 cells was detected by MTT assay. HBeAg and HBsAg expressions were measured by ELISA. HBV DNA copies were determined by real time quantitative PCR.RESULTS: A bulk of HMGN2 was isolated and purified from the acid soluble proteins of human lymph node, and identified. The HBV-transfected HepG2.2.15 cell line was used in the in vitro assay system.In the range of testing 1-100 mg/L of HMGN2, no cytotoxicity to HepG2.2.15 cells was detected by MTT assay.When incubated with HMGN2 at 1-5 mg/L for 72 h or 144 h, a significant reduction in HBeAg and HBsAg expression and in HBV DNA copies was observed in the supernatant of HepG2.2.15 cells. CONCLUSION: HMGN2 protein markedly inhibits HBV expression and replication in vitro.     

Role of renal preS1/S2-Ag and other HBV antigen determination in diagnosis of HBV-associated glomerulonephritis

AIM：To detect the expression of preS1/S2 antigen (preS1/S2-Ag) and other antigens of hepatitis B virus (HBV) in renal tissues of patients with HBV-associated glomerulonephritis (HBV-GN), and to analyze their roles in the diagnosis of HBV-GN.
METHODS：Patients hospitalized in our department from January in 2003 to January in 2013 were retrospectively studied. A total of 49 patients with positive HBV surface antigen (HBsAg) serology, clinical manifestations of hematuria and/or proteinuria, and pathological diagnosis of glomerulonephritis, without systemic lupus erythematosus, anaphylactic purpura, diabetes or hepatitis C, were selected. PreS1/S2-Ag, HBV e antigen (HBeAg), HBsAg and HBV core antigen (HBcAg) in the renal tissues were examined. Five cases of glomerular minimal-change disease (MCD) with negative HBsAg and 5 cases of non-glomerulonephritis with positive HBsAg served as controls. RESULTS：The positive rates of preS1/S2-Ag, HBeAg, HBsAg and HBcAg in the renal tissues from the 49 patients of glomerulonephritis with HBV infection were 32.7% (16 cases), 38.8% (19 cases), 14.3% (7 cases) and 46.9% (23 cases), respectively. Total antigen positive rate was 70.2% (36 cases). The expression of preS1/S2-Ag was located in the cytoplasm of renal tubular epithelial cells, glomerular epithelial cells, endothelial cells and mesangial cells, and positively correlated with the expression of HBcAg (r=0.459, P<001). The 4 antigens were not detected in the 5 cases of HBsAg-negative patients with glomerular MCD. In the 5 cases of HBsAg-positive patients with non-glomerulonephritis, there were 2 cases expressing HBeAg and 1 case expressing HBcAg, but no cases expressing preS1/S2-Ag or HBsAg. CONCLUSION：The expression of preS1/S2-Ag in renal tissues suggests that HBV may invade the cells of renal tissue. Combined detection of the 4 antigens could elevate the rate of diagnosis of HBV-GN.     

CD4+CD25+FOXP3+ Tregs and HBV-specific CTLs in peripheral blood from chronic hepatitis B patients

AIM: To investigate the roles of CD4⁺CD25⁺FOXP3⁺ regulatory T cells (Tregs) and HBV-specific cytotoxic T-lymphocytes (CTLs) in peripheral blood from the patients with chronic hepatitis B (CHB).METHODS: Peripheral blood mononuclear cells from 28 patients with CHB and 15 healthy controls were analyzed for Treg frequency using flow cytometry and for HBV-specific CTLs using enzyme-linked immunospot assay (ELISPOT).The clinical data of HBV-infected patients were considered.RESULTS: The frequency of CD4⁺CD25⁺FOXP3⁺Tregs was higher in the patients with CHB than that in the patients of healthy controls (3.14%±0.97% vs 1.95%±0.68%, P<0.05), and a positive correlation was found between Tregs and the DNA levels of HBV (r=0.831, P<0.01).HBV-specific CTLs were detected by ELISPOT in CHB patients and a negative correlation was observed between Tregs and CTLs (r=-0.540, P<0.01).CONCLUSION: Peripheral blood CD4⁺CD25⁺FOXP3⁺ Tregs in CHB patients are increased and closely correlated with the DNA replication of HBV and CTLs, suggesting that the clearance of HBV can be influenced by the inhibition of cellular immunoreaction through Tregs.     

Effects of xeroderma pigmentosum group D and p53 on replication of HBV

AIM:To investigate the effects of xeroderma pigmentosum D (XPD) and p53 on the replication of hepatitis B virus (HBV). METHODS:Recombinant plasmid pEGFP-N2/XPD and vacant vector plasmid pEGFP-N2 were transfected into HepG2.2.15 cells by liposome. On the next day, these cells were incubated with pifithrin-α, a p53 inhibitor, at a concentration of 20 μmol/L for 24 h. The cells were divided into 5 groups: blank control group, pEGFP-N2 group, pEGFP-N2/XPD group, pEGFP-N2/XPD+pifithrin-α group and pifithrin-α group. The mRNA expression of XPD, hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) and hepatitis B virus X protein (HBx) was detected by RT-PCR. The content of HBsAg and HBeAg in the supernatants of culture medium was measured by ELISA. The content of HBV-DNA in the supernatants of culture medium was examined by fluorescence quantitative PCR. Using the method of bDNA, the content of HBV-DNA in the core particles was assessed. RESULTS:The expression of XPD mRNA was elevated by the transfection of recombinant plasmid pEGFP-N2/XPD. The increase in XPD expression significantly down-regulated the mRNA expression of HBsAg, HBeAg and HBx. The content of HBsAg and HBeAg in the supernatants of culture medium was significantly decreased by the increase in XPD expression. The results of fluorescence quantitative PCR showed that the content of HBV-DNA in the supernatants of culture medium was significantly down-regulated by the increase in XPD expression. bDNA results showed that the content of HBV-DNA in the core particles was significantly decreased by the increase in XPD expression.Pifithrin-α abolished the above-mentioned effects of XPD (all P<0.01). CONCLUSION: XPD inhibits the replication of HBV through p53 pathway. Therefore, XPD and p53 may be the targets for antiviral therapy of hepatitis B.     

HTPP-MDC inhibits replication and subcellular localization of hepatitis B virus in HepG2.2.15 cells

AIM：To investigate the antiviral effect of hepatocyte-targeting and cell-penetrating peptide and Musca domestica cecropin (HTPP-MDC) fusion polypeptide against hepatitis B virus (HBV) in vitro, and to observe the penetrating ability of HTPP-MDC in hepatocytes. METHODS： HepG2.2.15 cells and Chang liver cells were co-cultured in vitro with HTPP-MDC at different concentrations. MTT assay was used to detect the cytotoxicity of HTPP-MDC in vitro. The levels of HBsAg, HBeAg and HBV DNA in HepG2.2.15 cell culture supernatants were measured by ELISA and real-time fluorescence quantitative PCR. The penetrating ability of HTPP-MDC was detected by laser confocal microscopy. RESULTS：The levels of HBsAg, HBeAg and HBV DNA in the supernatants of HepG2.2.15 cells treated with HTPP-MDC were remarkably reduced. The inhibitory effect of HTPP-MDC depended on the dose and action time of the drug. FITC-labeled HTPP-MDC was observed inside the cells under laser confocal microscope. CONCLUSION：HTPP-MDC strongly inhibits HBV replication in HepG2.2.15 cells. The penetrating ability of HTPP-MDC into hepatocytes indicates that HTPP-MDC is useful in clinic therapy for chronic hepatitis B-related diseases in the future.     

Two-unit ribozyme inhibit hepatitis B virus expressed in 2.2.15 cell

AIM: To observe the inhibition of HBc/HBeAg expression in the 2.2.15 cell transfected by two-unit ribozyme. METHODS: By use of subclone technique, two-unit ribozyme gene which was cutted from pGEM-Rz123 was ligated into the eukaryotic expression vector pBBS212. The recombinant plasmid was cotransfected into 2.2.15 cell using lipofectamine. Ribozyme was detected by dot-blot hybridization. The s and e/c antigen of HBV were detected by using ELISA, immunohistochemical technique, image analysis system and Western blot. RESULTS: After the transfected cell was selected two weeks by hygromycin B and G418. We found by dot-blot hybridization that ribozyme can express on 2.2.15 cell. HBeAg level can be reduced by 48.6% in the transfected 2.2.15 cell with two-ribozyme. Using immunohistochemical technique and image analysis system, Western blot, we observed that the level of HBcAg expressed in endocellular went down. CONCLUSION:Al these results strongly indicate that two-unit ribozyme can inhibit hepatitis B virus expression in the cell.     

Detection of miR-122 expression in serum by Taqman probe real-time fluorescence quantitative PCR and its clinical significance

AIM: To detect the serum level of miR-122 expression by the technique of Taqman probe real-time fluorescence quantitative PCR for identifying its clinical significance. METHODS: The stem-loop RT primer, PCR primer and Taqman probe of miR-122 and U6 snRNA were designed. The expression of miR-122 in the serum samples of 27 cases of preoperative hepatocellular carcinoma (HCC), 15 cases of hepatitis B, 15 cases of hepatitis C, 15 cases of healthy control (HC), 11 cases of postoperative hepatocellular carcinoma (PHCC) and 10 cases of recurrence after postoperative hepatocellular carcinoma was detected by Taqman probe real-time fluorescence quantitative PCR. U6 snRNA was used as the internal control. RESULTS: The results showed that the method of Taqman probe real-time fluorescence quantitative PCR could detect the amplification signal of serum miR-122. The expression level of serum miR-122 in the patients with HCC, hepatitis B, hepatitis C and recurrence was higher than that in HC and the patients with PHCC. Meanwhile, the serum level of miR-122 in the patients with hepatitis C was higher than that in the patients with HCC, hepatitis B and recurrence. However, no difference of miR-122 expression level among HCC, hepatitis B and recurrent patients was observed. The miR-122 level was lower in PHCC patients than that in HCC and recurrent patients. In hepatitis B virus surface antigen (HBsAg) and/or hepatitis B virus e antigen (HBeAg) positive patients, the miR-122 level was higher than that in the negative ones. The miR-122 level in hepatitis C antibody (HCV-Ab) positive patients was raised compared with the negative ones. The serum level of alanine aminotransferase (ALT) was positively correlated with the serum level of miR-122 (r=0.34, P<0.05). The miR-122 expression level in the patients with serum AFP≥400 μg/L was higher than that in the patients with serum AFP<400 μg/L. CONCLUSION: The method of Taqman probe real-time fluorescence quantitative PCR can detect the serum level of miR-122 expression. Serum miR-122 might be used as a new biomarker of liver diseases, especially in the early diagnosis of primary hepatocellular carcinoma, the curative effect of surgical operation and the prognosis.     

Changes of serum nitric oxide in CB4V-induced insulin-dependent diabetic mice

AIM: To observe the changes of serum nitric oxide and the production level of IL-1 in different period of coxsackievirus B4 (CB₄V)-induced insulin-dependent diabetic mice.METHODS: The insulin-dependent diabetes mellitus (IDDM) animal model induced by CB₄V infection was established. Serum nitric oxide level was estimated by nitrate reductase method after infection 72 h,1 week, 3 weeks,6 weeks,8 weeks, respectively. At the same time, level of IL-1 produced by peritoneal Mф was measued.RESULTS: (1) Changes of serum nitric oxide: serum nitric oxide level in control group remained normal level. The serum nitric oxide level in diabetic group increased significantly at 72 h after infection(P＜0.01), and reached top at 1 weeks after infection, then decreased to normal level in 6 and 8 weeks (P＞0.05). (2) IL-1 activities: IL-1 activities were increased obviously from 72 h to 3 weeks after virus infection, but decreased to normal level after 6 weeks.CONCLUSION: Nitric oxide may be one of the important factors in the development of CB₄V-induced IDDM.     

Association between IL-6-572C/G as well as IFNAR1-168G/C and hepatitis B virus infection in populations of Dai and Han ethnicities in Yunnan Province

ATM: To explore the association between IL-6-572C/G (rs1800796) as well as interferon alpha receptor 1 (IFNAR1)-168G/C (rs2257167) and prognosis after hepatitis B virus (HBV) infection in populations of Dai and Han ethnicities in Yunnan Province. METHODS: The blood samples were collected from Dai people and Han people, each nation including 100 healthy controls and 200 infected individuals (100 spontaneous recovery individuals and 100 chronic patients). Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing were used to identify the gene type. RESULTS: In Dai people, no significant difference was found between genetic polymorphism of -572C/G and prognosis after HBV infection. The differences of C and G alleles between spontaneous recovery group and chronic hepatitis B group, and healthy controls and HBV infection group were not statistically significant. Meanwhile, GG and CG genotypes were a vital protective factor for the person who developed into a chronic heptatitis B patient under the G allele dominance mode (GG+CG/CC) (P<0.05). In Han people, no statistically significance for IL-6-572C/G genotype and allele distribution in each group comparisons had been found, as well as the C allele recessive mode and C allele dominance mode. For the above 4 indicators, no statistically significant difference of IFNAR1-168C/G in Dai and Han people had been found.CONCLUSION: The GG+CG genotype of IL-6-572C/G may be a protective factor for the HBV-infected Dai people to develop into chronic hepatitis B patients. However, there is no significant association between the IFNAR1-168G/C polymorphism and prognosis after HBV infection in the 2 ethnicities.     

Study on mutations of exon BH of the androgen receptor gene in 45 cases of patients with prostate cancer

AIM and METHODS: To learn more about the mechanism of prostate cancer (PC) development and progression to androgen independence, the exons BH of the androgen receptor (AR) gene of forty-five patients with prostate cancer, six puncture tissues and thirty-nine slide tissues, were analyzed by polymerase chain reaction-single strand conformation technique (PCR-SSCP). RESULTS: Seven abnormal mobility shifts were found in five patients by PCR-SSCP. Combining the method with direct DNA cycle sequencing, two distinct missense (Glu872Gln, Met886Ile) point mutations were identified in puncture tissues from two patients of advanced prostate cancer with distant metastasis. These two point mutations represented two novel mutations. CONCLUSION: AR gene mutations might play an important role in the development and progression of prostate cancer.     

Seroprevalence of neutralizing antibodies to human adenovirus type 5, human adenovirus type 26 and chimpanzee adenovirus type 68 in patients with chronic hepatitis B and patients with primary liver cancer

AIM: To investigate the seroprevalence of neutralizing antibodies to human adenovirus type 5 (AdHu5), human adenovirus type 26 (AdHu26) and chimpanzee adenovirus type 68 (AdC68) in the patients with chronic hepatitis B (CHB) and the patients with primary liver cancer (PLC), and to provide guidance for developing safe and effective biotherapy vectors against CHB and PLC. METHODS: The blood samples from 196 patients with CHB and 193 patients with PLC were examined to assess the presence of neutralizing antibodies against AdHu5, AdHu26 and AdC68 by adenovirus neutralization assays. RESULTS: The seroprevalence rates of neutralizing antibodies to AdHu5, AdHu26 and AdC68 in the CHB patients were 84.7%, 58.2% and 39.8%, respectively. Among the patients with PLC, the prevalence rates of neutralizing antibodies were as follows: AdHu5, 75.1%; AdHu26, 66.8%; AdC68, 32.1%. CONCLUSION: The prevalence rates and titers of neutralizing antibodies against AdC68 were the lowest among the 3 adenoviruses. Therefore, AdC68 serves as more suitable biological therapy vectors for CHB and PLC than AdHu5 and AdHu26.     

Dynamic changes of circulating T FH cells during antiviral therapy in chronic hepatitis B patients

AIM:To investigate the circulating follicular helper T cells (T FH) and the expression of intracellular interleukin-21 (IL-21) before and after antiviral therapy in chronic hepatitis B (CHB) patients. METHODS:The CHB patients with antiviral therapy (14 cases of telbivudine therapy and 21 cases of interferon therapy) were selected in the study. Peripheral blood mononuclear cells (PBMC) were isolated by density gradient centrifugation. The level of CD4+CXCR5+ T FH cells and the expression of intracellular IL-21 were detected by flow cytometry. RESULTS:Intracellular IL-21 was produced mainly by T FH cells in peripheral blood T lymphocytes. The level of T FH cells had no statistically significant difference between the 2 different antiviral therapies in 24 weeks (P>0.05). The level of T FH cells and intracellular IL-21 in the patients with virological response (VR) was higher than those in the patients without virological response (NVR) before treatment. The levels of T FH cells was increased in NVR group during therapy. CONCLUSION:Antiviral therapy may contribute to elevate the level of circulating T FH cells in NVR patients. The high levels of circulating T FH cells and intracellular IL-21 before treatment can predict early virological response in CHB patients.     

Paired study on hepatitis B virus S gene mutation in immunoprophylaxis failure to prevent HBV vertical transmission

AIM：To explore the characteristics of hepatitis B virus S gene mutation in the vertical transmission after active and passive vaccination. METHODS：Fifteen cases of immunoprophylaxis failure were enrolled in the study. HBV S gene (including pres-S and S) from the mothers, newborns before active and passive vaccination and 7-month-old infants with immunoprophylaxis failure were detected by PCR amplification. The characteristics of HBV S gene mutation were compared among the 3 groups. RESULTS：The genotype of HBV in the newborns and the infants was the same as that in the mothers. The frequencies of mutation in the 2 fragments of the HBV S gene had no significant difference between the 3 groups. The homology tree model based on HBV S gene was analyzed in the 3 groups, in which every group had their own cluster. There were 15 different mutation sites between 7 pairs of mothers and newborns. There were 3 different mutation sites between 3 pairs of newborns and infants (nt273A→A/G, nt512C→C/T and nt1139C→A), among which the first 2 were located in the S gene region but not in the “a” determinant, and the latter was located in the overlap region of S and X genes. There were 25 different mutation sites between 9 pairs of mothers and infants, but only 1 case had a different mutation site between the mother, newborn and infant. CONCLUSION：The HBV species in newborns and infants with immunoprophylaxis failure were transmitted from the mothers. The mutations in the HBV S gene with immunoprophylaxis failure happened before and after active and passive vaccination, mainly before vaccination. The relationship between HBV S gene mutations and immunoprophylaxis failure should be further explored.