Quantitative analysis of HBV DNA level and HBeAg titer in hepatitis B surface antigen positive mothers and their babies: HBeAg passage through the placenta and the rate of decay in babies
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Abstract It is well documented that perinatal transmission is the major cause of chronic HBV infection in China. However, the mechanisms of HBV perinatal transmission are not defined clearly. It is not known whether hepatitis B e antigen can cross the human placenta, and the rate of HBeAg decay in babies with and without HBV breakthrough has not been studied. In this study, HBV serological markers were investigated in 95 hepatitis B surface antigen positive pregnant women. These markers were also studied in the babies at birth and at the age of 6 months and 12 months. The data show that 7.4% (7/95) children were infected with HBV during the first year after birth despite receiving passive‐active immunoprophylaxis with hepatitis B immune globulin and hepatitis B vaccine. The surface gene fragment of HBV DNA was cloned and sequenced following PCR amplification in 7 cases of HBsAg positive babies and their mothers. All babies had the same sequences as their mothers, although two babies also had sequences that would produce an amino acid substitution within the “a” determinant. Furthermore, we measured HBeAg titers and HBV DNA levels by using Abbott AxSYM system and LightCycler‐based real‐time fluorescence quantitative PCR in 54 mother‐infant pairs. Thirty‐three mothers were HBeAg positive, and 21 mothers were HBeAg negative. Seventy percent (23/33) of neonates from HBeAg‐positive mothers were HBeAg positive at birth compared with 0% (0/21) of neonates from HBeAg negative mothers. HBeAg was present at higher titer in the birth sera of the babies with HBV breakthrough than in babies without breakthrough. HBeAg was cleared from the serum in all 19 babies without breakthrough. In 17 of these 19 babies, the HBeAg was cleared within 6 months, and in two babies clearance took 12 months. The mean serum HBV DNA level in the mothers of the 4 infants with HBV breakthrough was significantly higher than in the mothers of babies who did not become infected. In conclusion, this data suggests that HBeAg can cross the human placenta, and disappears from serum within 6 months in most babies. HBV DNA levels in hepatitis B carrier mothers are associated with the failure of HBIG and vaccine immunization, and the additional influence of transmitted HBeAg cannot be excluded. J. Med. Virol. 71:360–366, 2003. © 2003 Wiley‐Liss, Inc.Keywords:
HBeAg
Hepatitis B
1000 pairs of maternal and cord blood samples were collected simultaneously at the time of delivery. 23 (2.3%) of the maternal samples were positive for HBsAg by enzyme-linked immunosorbent assay. HBeAg was detected in 11 (47%) of the 23 HBsAg positive mothers and anti-HBeAg was detected in another 5 samples. HBsAg and HBeAg were detected in 7 (30%) of the 23 cord blood samples from HBsAg-positive mothers, and anti-HBeAg was detected in one of these samples. At follow-up (6-18 months), antigenaemia had persisted in 17 (85%) of the 20 HBsAg-positive mothers and in 9 (45%) of 20 babies born to HBsAg-positive mothers. Seven of the 10 babies (70%) born to mothers positive for both HBsAg and HBeAg had persistent HBsAg in their blood, in contrast to 2 of the 10 babies (20%) born to mothers positive for HBsAg only. However, none of these mothers or their babies were found to have anti-HBeAg at follow-up. We conclude that the presence of HBeAg in mothers' blood enhances vertical transmission of hepatitis B virus infection to their babies.
HBeAg
Cord blood
Hepatitis B
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Objective: To study the inhibitory effect of Gantai capsules (肝泰胶囊, GTC) on replication and expression of HBsAg and HBeAg. Methods: The 2.2.15 cell line was chosen as a in vitro cell culture system, and by means of radioimmunoassay, the hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) in cell cultured medium were examined. Results: HBsAg, HBeAg values of 2.2.15 cells treated by GTC (200 μg/ml-1200 μg/ml) were lower than those of the control. And this inhibitory effect was in a dose dependent manner under experimental condition, GTC had no cytotoxicity. Conclusion: GTC could inhibit HBV replication and expression.
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Objective: To study the inhibitory effect of FFYC tablets (FFYC) on expression of HBeAg and HBsAg. Methods: The 2.2.15 cell line was cultured with cell cultured medium including different concentration of FFYC. By means of radioimmunoassay, the hepatitis B surface antigen ( HBsAg) and hepatitis B e antigen (HBeAg) were examined. Results: The max cytotoxicity concentration of FFYC was 600g/mL. Based on this concentration, FFYC was diluted and was added in cell culture medium. The results showed that HBeAg, HBsAg values of 2.2.15 cells treated by FFYC were lower than those in the control. Conclusion: FFYC could inhibit HBV replication and expression.
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Objective:To study genetic transmission from the parents with HBsAg to their children.Methods:Five serum HBV markers(HBsAg,anti HBs,HBeAg,anti HBe and anti HBc) and serum free HBV DNA were detected in 234 individuals from 117 families.Rusults:There was a significant difference in total infection rate of HBV,HBsAg,HBeAg,HBV DNA between the children of males with HBsAg and those of males with no HBsAg.The positive rate of HBV serum markers combination of father and their children was 52 4% in 63 families that father and children were all infected with HBsAg.HBV DNA mainly distributed in the combination that father and their children were all with HBsAg,there was a significant difference between that and other combinations.HBV DNA rised in accord with HBeAg.The childrens HBeAg and HBV DNA were all positive,who born after occurrence of HBeAg.HBV DNA of father with high titer HBeAg rised in accordence with there childrens.Conclusion:It showed that hepatitis B virus,can be transmitted by inheritance between the fathers and their children.
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Hepatitis B
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Hepatitis B virus (HBV) has been classified into 10 genotypes (A-I) and numerous subgenotypes (SGTs). There is growing evidence that HBV genotypes (GTs) influence clinical outcomes, HBeAg seroconversion rates, severity of liver disease, and response to interferon therapy. However, there is a paucity of data regarding their distinctive biological characteristics, in particular for GTF, the most prevalent in Latin America. To investigate the impact of HBV genotypes on HBV-DNA levels and viral antigen expression. Full-length HBV genomes representing GTs A-D and SGTs F1b and F4 were transfected in Huh7 cell line. Secreted HBeAg and intra and extracellular HBsAg were quantified by EIA. HBV-DNA was analyzed by real-time PCR. Marked differences were observed in HBV replicative capacity as well as HBeAg and HBsAg antigen expression across genotypes (Table 1). GTC secreted significantly higher levels of HBeAg in relation to the other GTs. GTD showed lower HBsAg extracellular levels than all other GTs, while GTA showed the highest HBsAg intracellular levels. Finally, SGTs F1b and F4 showed significantly lower HBV-DNA levels. Regarding the ratio of extra and intracellular HBsAg, GTs A and D showed the lower ratios compared to SGTs F1b and F4, while SGTs F1b and F4 showed the highest HBsAg/HBV-DNA ratio. This study provides new insights into the impact of HBV genotypes on HBV antigen expression and HBV-DNA levels. The uneven expression of antigens, as well as their intracellular accumulation, could be associated with the role of genotypes in pathogenesis. Likewise, the extracellular levels of HBsAg and the replication rate might have implications in immunopathogenesis as well as in the exhaustion of the host's immune system. The virus-cell interaction in different genotypes deserves further study to understand its role in the pathogenesis of HBV infection.
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Liver disease
Hepatitis B
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To constructed the shRNA expressing vectors targeting HBsAg gene and HBeAg gene of HBV, Pgs1, Pgs2, Pgs3 and psiHBV4, psiHBV6 in order to prove their inhibitation on the HBV antigen expression in outlive HepG2. 2. 15 cells.PTZ was used as negative control, both the shRNA expressing vectors targeting HBsAg and HBEAg gene of HBV was together transfected into HepG-2. 2. 15 cells with different combinations, and detected the expression liquid and the cultivated supernatant with MEIA after 24 h.The group transfecting psiHbv4 and PgS2, psiHBV4 and PgS3 could significantly inhibit HBsAg and HBeAg expression compared with control group (P < 0.05) in the cell disruption liquid and the cultivated supernatant. But the group transfecting psiHBV5 and PgS1, psiHBV6 and PgS3 cannot significantly inhibit HBeAg expression (P > 0.05) in the cell disruption liquid.The shRNA expressing vectors targeting HBsAg and HBeAg gene of HBV psiHBV4 and PgS2, psiHBV4 and HBeS3 could significantly inhibit the antigen expression of HBV than only one.
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Objective To study the effect of antisense phosphorothioate oligodeoxynucleotides (S-as ODN) of various regions of the HBV genome on the expression of HBsAg and HBeAg in the HepG22215 cell line. Method The expression levels of HBsAg and HBeAg in the HepG22215 cell line were determined by ELISA. According to gene structure of HBV, 5 S-as ODNs were designed and synthesized. The effect of different ASODNs on the expression of HBsAg and HBeAg was analyzed. Results This confirmed the characteristics of HepG22215 which could produce HBsAg and HBeAg. All 5 S-as ODNs could specifically inhibit the production of HBsAg and HBeAg in HepG22215 cells. A combination of drug and ASODN could enhance the inhibition. Conclusion These results suggest that S-as ODN may be a potential new therapy for anti-HBV. The inhibition showed sequence-specific, dose-dependent and drug-cooperative effects.
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OBJECTIVE To investigate the active parts of the extracts of the core of Litchi and their effect on the HBV. METHODS HepG 2.215 cell culture system was used for study of the expressions of HBsAg and HBeAg with A、B、C、D、E、F of the extract components from the core of litchi. RESULTS All extracts A、B、C、D、E 、F (200,100 mg·L -1) of the core of litchi. Were shown to inhibit the expression of HBsAg and HBeAg in HepG 2.2.15 cells, among which the extract E was the most effective, at 200 mg·L -1 in the medium, HBsAg expression was inhibited by 50%, and HBeAg by 20% in the third day of the experiment, and in the 9 th day, the HBsAg was inhibited by 90.9%, the HBeAg by 84.3%(P 0.01, compared to the control group). CONCLUSION The extracts of the core of litchi are very effective in inhibition of HBV in vitro.
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Objective To observe the effect of the serum containing Liuyueqing(LYQ)on the expression of HBsAg and HBeAg in HepG2.2.15 cells.Methods The toxicity of the serum containing LYQ on HepG2.2.15 cells was detected by means of MTT.The inhibitory effects of the serum containing LYQ on the excreation of HBsAg and HBeAg in HePG 2.2.15 cells were evaluated with method of enzyme linked immunosorbent assay(ELISA).Results The dose which the serum containing LYQ could inhibit 50% HepG2.2.15 cells was 11.56 g.(kg.d)-1.The dose which the serum containing LYQ could inhibit 50% HBsAg was 0.178g.(kg.d)-1,and its treatment index(TI)on HBsAg was 64.94.The inhibitory effect of the serum containing LYQ on HBeAg was not obvious.Conclusion The serum containing LYQ can inhibit HBV significantly in vitro,and it is a low toxic drug.
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MTT assay
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