脐带血乙肝标志物筛查乙肝病毒宫内感染的探讨

目的探讨脐带血乙肝标志物用于筛查乙肝病毒宫内感染的可能性。方法收集175例HBsAg阳性孕妇新生儿脐带血及24 h外周血,进行乙肝标志物和HBV DNA检测,6月龄时随访查外周血HBsAg。结果脐带血HBsAg阳性的新生儿外周血HBV DNA均为阳性,6月龄随访HBsAg阳性率均为100%。7例出生时注射了HBIG和乙肝疫苗的HBV DNA阳性的新生儿HBsAg随访阴转;8例脐带血HBsAg和HBV DNA均阴性的婴儿因直接母乳喂养HBsAg随访阳转。结论脐带血表面抗原可以作为筛查胎儿乙肝病毒宫内感染的指标。     

A randomized controlled clinical trial： Interruption of intrauterine transmission of hepatitis B virus infection with HBIG

AIM: To evaluate the efficacy of interruption of intrauterine infection of HBV with HBIG in pregnant women with positive HBeAg and HBsAg. METHODS: A prospective randomized controlled trial was adopted. Sixty cases with positive HBeAg and HBsAg were coincident with the criteria of inclusion, and 8 cases were excluded. Fifty-two cases were analyzed (28 cases in trial group and 24 in control group). All cases in trial group received 200 IU HBIG intravenously every 4 wk for 3 times from the 28th wk. The cases of control group received placebo in the same way. All pregnant women were detected for HBeAg and HBV-DNA at the beginning of the trial and end of the trial (delivery). The cord blood of all newborns were collected for detecting HBeAg and HBV-DNA simultaneously. RESULTS: For investigation of HBeAg of newborns in trial group, 6 of 28 cases of newborns had positive HBeAg, the HBeAg positive rate being 21.4%, the total rate of 95% CI being 8%-41%. In control group, 19 of 24 cases of newborns had positive HBeAg, HBeAg positive rate was 79.2%, the rate of 95%CI being 5%-93%. By statistical analysis, x2 = 17.26, P < 0.01, RR = 0.27, 95% CI (6.3×10-6, 8.6×10-5). For investigation of HBV-DNA of newborns in trial group, 7 of 28 cases of newborns had positive HBV-DNA, HBV-DNA positive rate being 25%, the total rate of 95% CI being ll%-45%. In control group, 20 of 24 cases of newborns had positive HBV-DNA, HBV-DNA positive rate was 83.3%, the total rate of 95% CI being 63%-95%. By statistical analysis, x2 = 17.62, P < 0.01, RR = 0.30, 95% CI (1.5×10-5, 1.7×10-4). The results indicated that there was significant difference in HBeAg positive rate and HBV-DNA positive rate of newborns between the two groups. In trial group, 7 of 28 newborns had HBV-DNA positive, but the HBV-DNA load of newborns was lower than that of their mothers. In control group, 20 of 24 newborns still had HBV-DNA positive, and the HBV-DNA load of newborns was close to those of their mothers. Statistical analysis indicated that there was no significant difference in HBV-DNA load between postnatal women without HBIG intervention and their filial generations (T = 81.5, P > 0.1). CONCLUSION: It is effective and safe to prevent in-trauterine infection of HBV with HBIG from the 28th wk in pregnant women with positive HBeAg and HBsAg. In clinical application, those pregnant women with negative HBeAg and positive HBV-DNA also need to be interrupted by HBIG.     

胎盘滋养层细胞的乙型肝炎病毒感染与宫内感染机制

目的：检测HBV对胎盘、胎肝和滋养层细胞的感染情况，探讨HBV的宫内感染机制。方法：研究对象包括20例孕妇胎盘组织、6例引产胎儿胎肝组织及体外培养的胎盘滋养层细胞。ELISA法检测孕妇外周血、胎儿脐血和6个月婴儿外周血HBV标志物；荧光定量PCR法检测血清和滋养层细胞中的HBV DNA；免疫组织化学法和免疫荧光法检测胎盘、胎肝组织及滋养层细胞中HBV标志物的表达；末端脱氧核糖核酸转移酶介导的缺口标记法（TUNEL）检测胎盘和滋养层细胞凋亡情况。结果：孕妇血清HBV DNA水平与胎儿脐血HBV DNA水平相关，脐血HBV DNA阳性者其母血HBV DNA〉1.0×10^7拷贝／mL；6例胎盘组织和3例引产胎儿胎肝组织中可见HBsAg免疫组织化学染色阳性细胞，其中1例胎肝组织中发现HBcAg阳性细胞；体外培养滋养层细胞与HBV DNA阳性血清共孵育后，可检测到HBsAg的表达，亦可检测到HBV DNA。体内和体外实验均检测到HBV感染后滋养层细胞凋亡呈增加趋势，且胎盘细胞的凋亡与脐血HBV DNA水平相关。体外实验结果显示，随感染时间的延长，滋养层细胞凋亡呈增加趋势。6个月后，12例新生儿有1例血清HBsAg、HBeAg和抗-HBc阳性，6例抗-HBs阳性。结论：HBV宫内感染的机制可能是通过HBV感染胎盘屏障而使胎儿发生HBV宫内感染。HBV在胎儿组织器官内的定位和复制可能是新生儿发生慢性HBV感染的重要因素。滋养层细胞凋亡可能是胎盘屏障阻断HBV宫内传播的一种保护性机制。     

Mechanism of intrauterine infection of hepatitis B virus

AIM:To explore the possible mechanism of intrauterine infection of hepatitis B virus (HBV).METHODS: HBV DNA was detected in vaginal secretion and amniotic fluid from 59 HBsAg-positive mothers and in venous blood of their newborns by PCR. HBsAg and HBcAg in placenta were determined by ABC immunohistochemistry.RESULTS:The rate of HBV intrauterine infection was 40.1% (24/59). HBV DNA was detected in 47.5% of amniotic fluid samples and 52.5% of vaginal secretion samples respectively.HBsAg and HBcAg were detected in placentas from HBsAg-positive mothers. The concentration of the two antigens decreased from the mother‘s side to the fetus‘s side, in the following order:maternal decidual cells&gt;trophoblastic cells&gt; villous mesenchymal cells&gt;villous capillary endothelial cells. However, in 4 placentas the distribution was in the reverse order. HBsAg and HBcAg were detected in amniotic epithelial cells from 32 mothers.CONCLUSION:The main route of HBV transmission from mother to fetus is transplacental, from the mother side of placenta to the fetus side. However, HBV intrauterine infection may take place through other routes.     

HBV宫内感染机制及基因的影响

乙型肝炎是一个世界性公共卫生问题。我国地处乙型肝炎高发区，人群中HBsAg阳性率高达10％以上。近年来的研究表明HBV宫内感染是HBV主要传播途径，亦是造成我国人群中大量乙肝病毒慢性感染者的主要原因，但对宫内感染机制尚无全面系统的研究。现将HBV宫内感染机制及易感因素的近期研究成果综述如下。     

乙型肝炎免疫球蛋白联合乙型肝炎疫苗阻断HBV母婴传播的效果分析

<正>HBV感染在世界范围内分布很广泛,其中我国HBV感染者高达9300万,约占总人口的7.18%[1]。据文献报道,约30%~50%HBV感染者是通过母婴传播所导致[2-3]。因此,切断HBV通过母婴垂直传播是当前疾控工作者重要且急迫的任务。根据濮阳市相关文件要求,自2012年开始,台前县HBs Ag阳性孕产妇所产新生儿均在出生后24 h内免费肌肉注射乙     

产前阻断对乙型肝炎病毒前C区和C启动子区的影响

目的 探讨产前阻断,即产前注射乙型肝炎免疫球蛋白(HBIG)对HBV前C区和基本C启动子(BCP)区核苷酸序列的影响.方法 无症状慢性HBV携带孕妇120例,产前阻断HBIG组67例,不产前阻断非HBIG组53例,荧光定量PCR法检测血清HBV DNA载量,ELISA检测血清HBsAg、HBeAg,套式PCR扩增HBV DNA前C区和BCP区核苷酸片段,DNA自动荧光测序仪对PCR产物直接测序.组间均数比较采用t检验,组间率比较采用卡方检验.结果 HBIG组33例孕妇收集到阻断前、后双份血清,其中23例阻断前、后HBV DNA前c区、BCP区均扩增测序成功.阻断前、后HBV前C区+BCP区、前C区、BCP区核苷酸的替代变异率分别为1.5%和1.4%、0.7%和0.6%、1.7%和1.7%(Fisher's精确检验,x2值分别为0.627、0.689、1.000,P＞0.05).阻断前、后1896G→A、1899G→A、1762A→T、1764G→A热点总变异发生率分别为27.2%和13.0%(x2=5.717,P=0.017),但各热点变异的发生率阻断前、后分别为30.4%和17.4%、17.4%和4.3%、26.1%和13.0%、34.8%和17.4%,差异无统计学意义(P＞0.05).扩增成功的53例HBIG组和47例非HBIG组孕妇临产时HBV DNA前C区+BCP区、前C区、BCP区核苷酸的替代变异率分别为0.9%和0.8%、0.3%和0.3%、1.1%和0.9%(Fisher's精确检验,x2值分别为0.434、0.839、0.340,P＞0.05);HBIG组和非HBIG组孕妇1896G→A、1899G→A、1762A→T、1764G→A热点总变异发生率及各热点变异发生率分别为5.7%和10.1%、9.4%和14.9%、0和2.1%、7.5%和10.6%、5.7%和12.8%,但差异无统计学意义(P＞0.05).结论 产前阻断可能不增加孕妇HBV DNA前C区、BCP区核苷酸变异;产前阻断可能减少孕妇体内HBV前C区、BCP区热点变异的发生.     

HBV宫内传播的研究进展

HBV宫内感染是乙肝免疫失败的主要原因, HBV宫内传播的途径主要有胎盘途径、PBMCS途径及经生殖细胞的传播. 其中胎盘的渗漏或胎盘细胞的感染是主要的途径. 母亲血清HBV DNA含量、HBV基因型、遗传因素是影响宫内感染的主要因素. 近年来的临床研究显示孕期使用乙肝高效价免疫球蛋白或拉米夫定可以阻断大部分宫内感染, 但目前尚缺乏大样本的随机对照研究, 探索宫内感染的机制和可行的阻断措施是我国控制乙型肝炎流行的关键所在.     

Molecular characterization of occult and overt hepatitis B (HBV) infection in an HIV-infected person with reactivation of HBV after antiretroviral treatment interruption

Introduction  

Occult HBV infection is characterized by the absence of surface antigenemia and the presence of potentially infectious hepatitis B virus (HBV)-DNA present in liver, serum, or both. Reactivation of chronic HBV infection in the presence of the HBV surface antigen (HBsAg) is a well-known complication in immunocompromised individuals under cytotoxic chemotherapy or in HIV-infected individuals when nucleos(t)ide analogs effective against HIV/HBV are discontinued. However, little is known on the possibility of such a complication in HIV-infected persons with HBV-core antibody (anti-HBc) as the sole serological marker of past HBV infection.     

乙型肝炎病毒宫内感染研究现状

乙型肝炎(乙肝)病毒(hepatitis B virus,HBV)宫内感染是我国HBV携带者和慢性乙型肝炎(慢乙肝)发生的重要原因。胎儿期的HBV感染不仅易形成以后的慢性携带状态,而且是肝硬化和肝癌的高危因素。对HBV宫内感染的研究主要集中在国     

Hepatitis B immunoglobulin (HBIG) efficacy in the prevention of perinatal transmission of hepatitis B virus (HBV) infection

Twelve out of 13 infants born from mothers having both HBsAg and HBeAg developed HBV carrier state within 4 months after birth. On the other hand, no babies from mothers having HBsAg and anti-HBe developed HBV carrier state. In order to prevent perinatal transmission of HBV, HBIG was administered into 14 babies born from mothers with positive HBeAg three or four times during 6 months after birth. During 12 months or more of observation period 5 out of 14 infants who received HBIG acquired active anti-HBs response after discontinuation of HBIG (passive-active immunization). However, 3 out of 14 infants unfortunately developed persistently positive HBsAg antigenemia at 12th, 14th and 14th month respectively after birth. Remaining 6 babies still have no virus markers, indicating not infected. These results indicates that HBIG administration was extremely effective for prevention of perinatal transmission of HBV. However, additional preventive measures with active immunization (HBV vaccine) seems to be necessary to prevent completely the perinatal transmission of HBV.     

Glomerulonephropathy associated with hepatitis B virus (HBV) infection

Forty-seven renal biopsies of glomerulonephropathy with persistent Australian antigenaemia (HBsAg is mostly positive) were studied with light microscope, electron microscope and direct immunofluorescence. Immunohistochemical method (ABC method) was used to examine HBsAg, HBeAg and HBcAg deposits in renal tissue. In addition 20 cases of idiopathic membranous nephropathy (MN) were studied for comparison. These 47 cases included 19 children and 28 adults. The results indicated that Australian antigens diffusely deposited in glomeruli in 14 cases (29.7%), with HBsAg in 7 cases (50.0%), HBeAg in 13 cases (92.8%) and HBcAg in 2 cases (14.3%). The 14 positive cases included 11 children and 3 adults. The pathologic types were membranous nephropathy in 12 and membranoproliferative glomerulonephritis in 2 cases. The membranous type was characterized by irregular thickening of capillary wall and double contour, bubble-like appearance and spike formation of glomerular basement membrane (GBM); immune complexes and electron dense deposits may be present in different sites of glomeruli. Coarse granular deposits of IgG and C3 along GBM were the principal pattern, but IgA, IgM and C1q were often present. Among the 20 idiopathic MN, 2 were found to have HBeAg deposition along GBM, one was a child and the other an adult IgG, IgA, IgM, C3 and C1q with HBeAg deposits were present in glomeruli.     

Chronic hepatitis B virus (HBV) infection in pregnancy

Background  Hepatitis B is a considerable disease burden among Asians. Little is known about its disease behaviour in pregnant women. Methods  Clinical, laboratory and radiological data of pregnant and peri-partum females with chronic hepatitis B virus (HBV) infection who were seen between years 1999 and 2004 were studied. Their progress was documented up to 6 months post-partum. This was compared with the age-matched and HBe status-matched, non-pregnant, female patients with chronic HBV infection, who were consecutively selected from the department’s registry as controls (ratio 1 mother: 4 non-pregnant controls), over the corresponding period. Results  A total of 35 mothers and 140 controls were studied. Mean age of patients was 30.7 ± 3.6 years. Majority of mothers (74.3%) presented during pregnancy itself. 1st:2nd:3rd trimester presentation = 20.0%:48.6%:5.7%. Majority (65.7%) were positive for HBe antigen (HBeAg) at the time of presentation. About 57.1% mothers had a clinical event in the form of alanine transferase (ALT) elevation and/or loss of HBeAg vs 28.8% among controls (P = 0.002). Among HBeAg-positive subjects, more mothers (14.3%) than controls (2.2%) had resultant HBeAg loss (P = 0.02). Among HBeAg negative subjects, more mothers than controls had serum ALT elevations in the post-partum period (P = 0.007). Overall, more mothers had elevated ALT levels than controls, regardless of their HBeAg status. Neither mothers nor control subjects decompensated clinically, neither required liver transplantation nor died during the study period. Conclusions  Pregnancy is associated with serum ALT elevation and HBeAg loss in patients with chronic HBV infection in the peri-partum period.     

Effect of HIV infection and antiretroviral therapy on hepatitis B virus (HBV)-specific T cell responses in patients who have resolved HBV infection

Coinfection with hepatitis B virus (HBV) is a common occurrence in human immunodeficiency virus (HIV)-positive patients and an increasing cause of morbidity and mortality. The CD8(+) T cell response is critical for long-term control of HBV in patients resolving acute infection. Here, we examine the effect of HIV on HBV-specific CD8(+) T cell responses in patients who have resolved HBV infection. A cross-sectional study showed a reduction in HBV-specific CD8(+) T cell responses in HIV-positive, HBV-immune patients, compared with those in HIV-negative, HBV-immune patients. A longitudinal study of a subgroup of patients examined whether this attrition could be reversed by effective antiretroviral therapy. The introduction of highly active antiretroviral therapy (HAART) resulted in reconstitution of some HBV-specific CD4(+) and CD8(+) T cell responses, in association with restoration of CD4(+) T cell counts. These data provide a mechanism to account for the observed impairment of control of HBV infection in the setting of HIV infection and support the ability of HAART to reconstitute functionally active T cell responses.     

Genomic heterogeneity of hepatitis B virus (HBV) and outcome of perinatal HBV infection

BACKGROUND/AIMS: Data regarding hepatitis B virus (HBV) genomic heterogeneity in perinatal infection are incomplete, although HBV variants might be involved in neonatal fulminant hepatitis (ALF). We investigated HBV variability in infected babies showing different clinical courses. METHODS: We analyzed HBV genomes isolated from nine vertically infected babies and the mothers of four of them. Two infants born to HBe-antigen (HBeAg)-positive women developed a chronic infection; seven babies (six born to anti-HBe mothers) developed acute hepatitis that had a fulminant course in four cases and a benign course in three. Two babies developing ALF received anti-HBV immunoprophylaxis at birth. RESULTS: Viruses carrying no significant mutation infected infants born to HBeAg-positive women. HBeAg-defective viruses were detected both in children with benign and fulminant hepatitis and their mothers. A double nucleotide mutation at positions 1762 and 1764 of the HBV core-promoter was found in two of the four infants with ALF, although it was not detected in isolates from the mother of one of them. No significant S gene mutation was found in HBV from any of the babies. CONCLUSIONS: This study indicates that HBV genomic heterogeneity is not primarily involved either in the evolution of the infection or the failure of neonatal HBV immunoprophylaxis.     

慢性乙型肝炎患者体内乙型肝炎病毒准种特点的初步研究

目的 探讨乙型肝炎病毒（HBV）准种在慢性乙型肝炎患者中的存在状态。方法 以已知中国株HBV基因序列为依据。设计特异性聚合酶链反应（PCR）引物,自3例慢性乙型肝炎患者体内扩增HBV全S基因片段,克隆人pGEMTeasy质粒,用限制性片段长度多态性（RFLP）法确定HBV的变异现象,DNA测序确定病毒的变异程度。结果 分别自3例患者血清中克隆出29、28和8个阳性克隆,以EcoRⅠ酶切患者1和2的RFLP结果提示各有5种不同的长度带型,PCR产物XhoI酶切RFLP结果表现为高度保守,测序结果发现HBV体内毒株DNA序列高度一致,同一患者两株全S区序列测序结果表明DNA序列的同源性大于97%。结论 慢性乙型肝炎患者体内有HBV准种共存,且呈现出一定的优势克隆现象。可能与患者预后有关。