Recombinant vaccinia vector-based vaccine (Tiantan) boosting a novel HBV subunit vaccine induced more robust and lasting immunity in rhesus macaques

This study explored several prime-boost strategies in rhesus macaques using various novel hepatitis B virus (HBV) vaccines that showed promise as prophylactic and therapeutic approaches in our previous study using in a mouse model. The tested vaccines included an HBV particle subunit (HBSS1) vaccine and the recombinant vaccinia (RVJSS1) or adenoviral (rAdSS1) vector-based vaccines containing S (1–223 aa) and PreS1 (21–47 aa). The strength and maintenance of humoral activity (IgG and neutralizing antibodies) and cellular immunity (interferon-γ production assessed by IFN-γ enzyme-linked immunosorbent spot (ELISpot) assay) were investigated in a longitudinal study following various vaccination protocols until 79 weeks post-vaccination. We found that HBSS1/RVJSS1 heterologous prime-boost elicits similar strong humoral immunity but more robust and lasting cellular immunity (CMI) than HBSS1/HBSS1 homologous vaccination in rhesus macaques. Furthermore, HBSS1/RVJSS1/RVJSS1 induced more robust and lasting CMI in macaques than did HBSS1/HBSS1/rAdSS1 vaccination. Therefore, HBSS1/RVJSS1/RVJSS1 is most promising candidates for protecting humans against HBV infection, especially for therapeutic application.     

Immunity of two novel hepatitis C virus polyepitope vaccines

Hepatitis C virus (HCV) infection remains a serious public health burden around the world. So far there is no effective vaccine against this virus. Neutralizing antibody (NAb) responses to the epitopes within HCV E1 and E2 proteins are related to the resolution of hepatitis C infection. E. coli heat-labile enterotoxin B subunit (LTB) has been described as potent immunity adjuvants. In this study, we constructed recombinant pET vectors: pET-R9-Bp (B cell polyepitopes) expressing 7 epitopes from HCV E1 and E2 proteins including R9 (E2_384-411aa)-Bp (E1_313-327aa-E2_396-424aa-E2_436-447aa-E2_523-540aa-E2_610-627aa-E2_631-648aa) and pET-LTB-R9-Bp expressing LTB adjuvant in combination with R9-Bp. Recombinant proteins R9-Bp and LTB-R9-Bp were expressed successfully in E. coli and purified by the Ni-NTA column. Both R9-Bp and LTB-R9-Bp in BALB/c mice induced robust humoral immune response in the context of intraperitoneal or intramuscular immunization but not oral immunization. Intraperitoneal administration of LTB-R9-Bp induced a higher antibody titer (peak titer: 1:341000) than that of R9-Bp (peak titer: 1:85000) after the second boost (P = 0.0036 or 0.0002). However, comparable antibody peak titers were elicited for both R9-Bp and LTB-R9-Bp in intramuscular immunization albeit with significant difference (P = 0.0032) a week after the second boost. In addition, both R9-Bp and LTB-R9-Bp induced the secretion of cytokines including IFN-γ and IL-4 at similar levels. anti-sera induced by both R9-Bp and LTB-R9-Bp recognized native HCV E1 and E2 proteins. Moreover, these HCV-specific antisera inhibited significantly the entry of HCV (P < 0.0001). Taken together, these findings showed that E. coli-based both R9-Bp and LTB-R9-Bp could become promising HCV vaccines.     

Development of a more efficient hepatitis B virus vaccine by targeting hepatitis B virus preS to dendritic cells

BackgroundConventional hepatitis B virus (HBV) vaccines fail to induce protective antibody titers in 5–10% of immune-competent vaccinees. Therefore, there remains an urgent need to develop a safe and effective HBV vaccine.MethodsIn this study, we developed an effective and economical method for producing the HBV vaccine by using the high binding capacity of biotin–streptavidin. The preS antigen of HBV was fused with the core streptavidin (cSA) moiety (preS-cSA) and highly expressed in Escherichia coli. We investigated whether the preS-cSA protein could target dendritic cells (DCs) by binding a biotinylated antibody against the DC receptor CD205 (biotin-αCD205). Moreover, we evaluated the preS-cSA/biotin-αCD205 complex as a candidate vaccine by detecting the humoral and cellular immune responses elicited by this vaccine.ResultsOur data show that the preS-cSA/biotin-αCD205 complex targeted DCs and induced high anti-HBV antibody titers of IgG2a, IgG1, and IgG in vivo. Furthermore, vaccination with the preS-cSA/biotin-αCD205 complex prevented HBV infection in female mice. More interestingly, this novel vaccine exerted a therapeutic role in mice with HBV infection.ConclusionsTaken together, our results reveal that the preS-cSA/biotin-αCD205 vaccine induces effective immunological protection against HBV, and is a promising candidate for preventing HBV infections.     

乙型肝炎疫苗长期免疫对人群乙型肝炎病毒流行状况的影响

目的探讨乙型肝炎（乙肝）疫苗长期免疫接种后整体人群乙肝病毒（HBV）感染状况及其变化趋势。方法采用整群抽样结合横断面调查方法，共收集资料完整的调查对象4686名，采集静脉血并分离血清，用固相放射免疫法检测HBV感染标志。结果整体人群平均HBsAg阳性率为7．5％，抗-HBs为44．5％，抗-HBc为47．8％；0～19岁人群HBsAg和抗-HBc阳性率较≥20岁人群显著下降。乙肝疫苗免疫组的HBsAg阳性率为2．8％，抗-HBc阳性率为12．0％，HBV感染率为12．5％，未免疫组分别为10．2％、69．8％和71．2％。男性平均HBsAg阳性率比女性高，抗-HBc和抗-HBs阳性率男女性别间无差异。0～19岁人群的HBsAg阳性率为2．4％，而20～30岁人群阳性率达到13．6％～17．7％，到60岁开始下降；0～19岁人群的抗-HBs阳性率随年龄增长而明显下降，≥20岁人群的抗-HBs、抗-HBc阳性率均随着年龄增长而呈升高趋势。结论长期开展新生儿乙肝疫苗免疫使人群HBV流行状况发生变化，感染高峰年龄段后移。     

Duration of hepatitis B antibody response in children immunised with hepatitis B and compulsory vaccines

Twenty four subjects were simultaneously administered DT toxoids, OPV and HBV vaccines at the age of 3, 4–5 and 11 months and then followed up for 2 and 4 years in order to evaluate the duration of the immune response and the need and the timing of HBV revaccination. A fall in anti-HBs titre below 10 mIU/ml was observed at the follow up in 4/24 (16.7%) of the subjects. In other 5 children (20.8%) anti-HBs titre was found to be just above 10 mIU/ml. This would suggest that a revaccination is indicated and it could be performed at the age of 5–6 years when children enter school. This schedule is simple, effective and money saving since it reduces the cost/benefit ratio and the number of visits for immunisations, and it is expected to improve the compliance for the vaccination.     

联合免疫对乙型肝炎表面抗原阳性的慢性乙型肝炎病毒感染孕妇子女的免疫效果

目的 评价乙型肝炎疫苗（HepB）联合乙型肝炎免疫球蛋白（HBIG）对乙型肝炎表面抗原（HBsAg）阳性的慢性乙型肝炎病毒（HBV）感染孕妇子女的免疫效果.方法 326例乙型肝炎e抗原（HBeAg）阴性HBV感染孕妇及其分娩的375例子女纳入本研究,记录母亲孕期HBIG使用情况、分娩方式、子女出生后免疫预防措施和喂养方式,并进行HBV标志物比较.结果 375例子女均在出生12h内使用了HBIG,352例（93.9％）出生24 h内接种了第1针HepB,23例（6.1％）均因存在各种新生儿疾病而延迟,但分别在出生后7 ～42 d进行补接种.236例行脐带血检测HBV标志物,39例（16.5％）HBsAg阳性,197例（83.5％）HBsAg阴性,脐带血HBsAg阳性与阴性子女抗-HBs阳性率及抗-HBs中位浓度比较差异无统计学意义（P＞0.05）.孕期使用HBIG与未使用HBIG母亲的子女抗-HBs阳性率分别为63.5％（47/74）,59.8％（180/301）;剖宫产与自然分娩的子女抗-HBs阳性率分别为65.2％（103/158）,57.1％（124/217）,母乳喂养、混合喂养、人工喂养的子女抗-HBs阳性率分别为62.0％（119/192）,58.9％（63/107）,59.2％（45/76）,差异均无统计学意义（P＞0.05）.结论 HBeAg阴性HBV感染孕妇的子女经正规的HBIG联合HepB免疫预防后,不同的分娩方式、喂养方式、孕晚期是否使用HBIG对子女HepB的免疫应答与HBV的母婴传播无影响.     

HBsAg阳性孕产妇其幼儿乙型肝炎疫苗免疫应答情况及其影响因素

目的  获取HBsAg阳性孕产妇其幼儿乙型肝炎(乙肝)疫苗无/弱应答的概率,分析其影响因素。 方法  对2016年1月至2017年4月在陕西省西北妇女儿童医院分娩的284对HBsAg阳性孕产妇及其幼儿进行研究,随访幼儿乙肝疫苗接种情况及血清学标志物产生情况。 结果  高危幼儿无/弱应答率为10.57%(28/237)。表面抗原(HBsAg)、e抗原(HBeAg)和核心抗体(HBcAb)均阳性的孕产妇其幼儿发生乙肝疫苗免疫无/弱应答率为19.64%,高于HBsAg、e抗体(HBeAb)和HBcAb均阳性的孕产妇的幼儿,其乙肝免疫无/弱应答率为9.89%(RR=1.99,95% CI：1.01~3.92,P＜0.001)。孕期有穿刺史者其幼儿发生无/弱应答的风险是无穿刺史者的6.72倍(RR=6.72,95% CI：2.79~16.19,P=0.049),幼儿发生乙肝疫苗无/弱应答的孕产妇白蛋白(ALB)低于强应答组(t=2.518,P=0.013),白球比(A/G)高于强应答组(t=-5.559,P＜0.001)。 结论  HBsAg阳性孕产妇幼儿是乙肝疫苗无/弱应答的重点人群,其又处于母亲为传染源的高危环境中,应重点进行抗体监测。其孕期有创检查可能会增加幼儿发生乙肝疫苗无/弱应答的概率。HBsAg、HBeAg和HBcAb均表现为阳性的孕产妇幼儿有较高的乙肝疫苗免疫无/弱应答率。     

Toll样受体抗乙型肝炎病毒感染研究

Toll样受体(TLR)是近年来备受瞩目的一种跨膜模式识别受体。作为天然免疫的重要组分之一,TLR通过配体识别、信号转导、免疫分子活化等环节发挥强大的防御功能;同时也在联系天然免疫和固有免疫方面发挥重要作用。此文就近年来TLR在HBV感染和相关免疫调节机制的研究及其在临床上的应用进行综述。     

核苷类抗病毒治疗对慢性乙型肝炎相关性肝癌患者术后预后的影响

目的研究慢性乙型肝炎(慢乙肝)患者外周血微小RNA(miR)-155水平及其与免疫活化的关系。方法收集81例慢乙肝患者、77例HBV携带者和51例正常对照者。检测肝功能、HBV DNA和乙肝三系,用流式细胞术检测外周血CD₄⁺和CD₈⁺T细胞水平及其活化水平(CD₃₈⁺比例)。免疫磁珠分选外周血CD₄⁺和CD₈⁺T细胞,提取总RNA,实时定量PCR法检测CD₄⁺和CD₈⁺T细胞中相对miR-155水平。结果慢乙肝组外周血CD₄⁺和CD₈⁺T细胞中相对miR-155水平分别为689.37±139.43和2735.05±397.56,高于HBV携带组(420.64±101.32和1287.46±323.51)和对照组(271.83±72.95和624.62±158.33)的水平(P＜0.05),HBV携带组CD₈⁺T细胞中相对miR-155水平高于对照组(P＜0.01)。慢乙肝组CD₄⁺和CD₈⁺T细胞活化水平分别为(52.93±8.64)%和(69.28±14.85)%,高于对照组[(22.34±5.98)%和(28.64±9.53)%](P＜0.01),并且与miR-155水平呈正相关。结论慢乙肝患者外周血CD₄⁺和CD₈⁺T细胞中相对miR-155水平显著升高,可能与免疫活化呈正相关。     

乙型肝炎病毒e抗原结构、功能及变异的临床意义

HBeAg并非HBV装配、复制或感染所必需,但却具有免疫调节和影响病毒复制的重要功能,而HBeAg变异后出现的HBeAg阴性慢性乙型肝炎更有特殊的临床特征。此文就近年有关HBeAg结构及功能的研究进展及其变异临床意义进行综述。     

重组乙型肝炎疫苗阻断乙型肝炎病毒母婴传播方案的保护效果评价

目的 评价国产重组乙型肝炎(乙肝)疫苗及其与中效价乙肝免疫球蛋白(HBIG)联合应用母婴阻断方案的保护效果。方法 在广西等3个地区，对乙肝病毒双阳性母亲新生儿，应用重组乙肝疫苗和重组乙肝疫苗加50IU HBIG两种母婴阻断方案免疫新生儿，共随访单纯重组乙肝疫苗母婴阻断儿289例，重组乙肝疫苗加HBIG阻断儿186例。结果 单纯重组乙肝疫苗的母婴阻断效果为87．8％(95％CI：83．6—91．9)，重组乙肝疫苗加HBIG的阻断效果为91．2％(95％CI：86．7—95．6)，重组(酵母)乙肝疫苗和重组(CHO细胞)乙肝疫苗间(P=0．7072)、两种母婴阻断方案间(P=0．2955)及各地免疫人群间(P=0．9987)的母婴阻断效果差异均无显著统计学意义。两种母婴阻断方案免疫新生儿间抗—HBs的阳转率(P＝0．3188)和抗体滴度(GMT)间(P＝0．8925)差异均无显著统计学意义，首剂免疫后1年，抗—HBs阳性率在单纯重组乙肝疫苗组和重组乙肝疫苗加HBIG组分别为91．1％和93．5％，GMT分别为153mIU／ml和164mIU／ml，以后逐年显著下降(线性趋势检验χ^2＝60．47，P=0．0001)，至免疫后第4年，阳性率分别降为65．0％和66．6％，GMT仅为第一年的1／3。结论 重组乙肝疫苗加中效价HBIG的母婴阻断效果可达90％以上，明显优于常规剂量的血源疫苗。中国现行重组乙肝疫苗抗—HBs的免疫检测技术方法有待改进。     

乙型肝炎病毒感染免疫耐受机制的研究进展

HBV感染后容易形成慢性化,多数学者认为其主要机制是宿主对HBV各种抗原产生免疫耐受。宿主在对HBV免疫应答过程中的各个环节都可能形成免疫耐受。此文就近年来慢性HBV感染的患者体内免疫耐受机制的研究进展进行综述。     

无锡市城区20岁以上人群乙型肝炎病毒感染及免疫状况调查

目的 对江苏省无锡市城区20岁以上自然人群HBV感染与乙型肝炎(乙肝)疫苗接种关系进行研究.方法 按知情、自愿、随机的原则抽取无锡市20岁以上的自然人群3744名进行乙肝血清流行率和乙肝疫苗接种调查,采用ELISA和放射免疫分析法测定乙肝五项指标(HBsAg、抗-HBS、HBeAg、抗-HBe和抗-HBc).结果 3744名调查对象总HBV感染率经标化后为51.7%,HBsAg、抗-HBs、HBeAg、抗-HBe、抗-HBc阳性率经标化后分别为4.5%、48.5%、0.3%、3.5%、51.4%.30岁以下人群HBsAg阳性率最低,分别为2.9%和2.6%.抗-HBc阳性率男性显著高于女性(P<0.05),且随着年龄的增加而逐步升高(趋势χ2=256.2,P<0.001).该人群乙肝疫苗标化接种率为17.6%,随着年龄的增加接种率迅速下降(P<0.05).接种疫苗人群的HBsAg阳性率和HBV感染率明显低于未接种疫苗人群、抗-HBs阳性率则高于未接种人群(P值均<0.05).结论 成年人接种乙肝疫苗可以影响整个人群的HBV感染模式,在加强新生儿乙肝疫苗计划免疫同时,应进一步加强对成年人的乙肝疫苗免疫策略.     

B7家族共刺激分子与机体对乙型肝炎病毒感染的免疫耐受

机体对HBV的免疫耐受是HBV感染慢性化的主要机制.共刺激分子是控制免疫应答的开关.B7家族共刺激分子是最基本的共刺激分子,在HBV感染中可提供细胞活化所需要的第二信号,决定T细胞是活化增殖还是转为无能状态,对机体的免疫耐受起重要作用.了解B7家族共刺激分子在HBV感染免疫应答中的作用,对揭示HBV感染免疫耐受机制有重要的意义.     

母亲乙型肝炎病毒携带状况与新生儿乙肝疫苗免疫效果关系的探讨

目的分析母亲乙肝病毒（hepatitis B virus,HBV）携带状况与新生儿乙肝疫苗（hepatitis B vaccine,HepB）免疫效果之间的关系,为阻断HBV母婴传播提供依据。方法在邯郸市随机抽取8022名满7月龄--2周岁儿童及其母亲,调查儿童的HepB接种情况,采取试纸和酶联免疫吸附试验方法检测母子HBV标志物携带状况,并检测HBsAg阳性母亲的HBV基因型。结果母亲HBsAg阳性率为2．43％,儿童阳性率为0．45％,HepB保护率为81．48％。143名HBsAg阳性母亲中有127名C型,占97．69％。26对母子名HBsAg均阳性的母亲中,大三阳9名,小三阳5名,HBsAg和HBcAb阳性10名,三者差异有统计学意义（χ^2=6．03,P〈0．05）。结论母亲HBV基因型中以C型为主。阻断HBsAg阳性的母婴传播应采取联合接种HepB和乙肝高效免疫球蛋白等综合措施,同时,HBsAg和HBcAb双阳性母亲在母婴传播中的意义不可忽视。     

乙型肝炎免疫球蛋白阻断乙肝病毒母婴传播过程中病毒S区基因变异的研究

目的了解乙型肝炎免疫球蛋白（HBIG）在阻断母婴传播过程中S基因变异的特点,比较与未予阻断者的差异,探讨基因变异与宫内传播的关系,协助评价HBIG的治疗安全性.方法将18对乙型肝炎病毒（HBV）携带母亲及其宫内感染新生儿按母亲产前处理分为HBIG组8对,母亲于产前3月起肌肉注射HBIG 200～400IU至分娩前;对照组10对,母亲产前不接受HBIG者.巢式PCR扩增HBV S基因片段并直接测序.以每对病例第一份（母亲孕中期）血清HBV株S基因区氨基酸（或核苷酸）作为标准,对每对病例第二份血清（母亲分娩前）和第三份血清（新生儿）HBV株进行分析.结果HBV S区碱基替代突变率和氨基酸变异数在HBIG组和对照组之间的差异均无统计学意义;18例宫内感染儿17例其病毒株与母亲分娩前的优势株一致,其中4例为S区变异株;18例宫内感染儿病毒株分型B型adw2 12例,C型adrq^＋6例.结论无症状携带HBV孕妇产前使用现有剂量HBIG至临产并未增加HBV S区的变异.HBV S区变异株和未变异株均可经宫内传播,宫内感染发生于孕后期;HBV S区变异并非发生宫内感染的主要原因.     

乙肝病毒前S1抗原检测在入境人员检测诊断中的应用价值

目的评价乙肝病毒PreS1抗原在入境人员检测中的应用价值。方法将2008—2010年经我中心体检的362份入境乙肝病毒表面抗原(HBsAg)阳性的血清标本进行PreS1抗原检测,同时用PCR法进行乙型肝炎病毒基因(HBV-DNA)测定。结果PreS1总阳性率为41.16%,HBV-DNA阳性率为43.92%,乙肝病毒e抗原(HBeAg)阳性率为29.83%。PreS1总阳性率明显高于HBeAg阳性率。PreS1与HBV-DNA的总符合率为84.80%。结论PreSl抗原作为乙肝感染和复制的血清学指标敏感性高于HBeAg,对提示乙肝患者的病情发展和转归有重要临床意义。PreS1抗原可以作为一项新的乙肝病毒复制的指标在入境人员中进行筛查。     

Persistence of antibodies and immune memory to hepatitis B vaccine 20 years after infant vaccination in Thailand

Booster vaccination against hepatitis B (HBV) is not currently recommended, although debate continues on the duration of protection after priming. We assessed antibody persistence and immune memory to hepatitis B 20 years after priming with a recombinant HBV-vaccine during infancy. Infants were vaccinated at birth, 1, 2 and 12 months of age. A subset received a booster dose at Year 5. Antibody persistence was measured approximately yearly until Year 20. Immune memory was assessed by administration of HBV booster dose. At Year 20, anti-HBs seroprotection rates and GMCs tended to be higher in Year 5 boosted than unboosted recipients (83.9% versus 60.5%). After the Year 20 booster dose, anti-HBs anamnestic responses were within the same range 95.8% of subjects in both groups. Primary and booster vaccination with HBV-vaccine in infants induces sustained seroprotection and immune memory against hepatitis B for up to 20 years. Higher persisting seroprotection rates in subjects boosted at Year 5 did not translate into apparent differences in immune memory in a high endemic country.     

Minimization of hepatitis B infection among children in Jiangsu,China, 12 years after integration of hepatitis B vaccine into the expanded program on immunization

BackgroundChina has integrated hepatitis B vaccine into the Expanded Program on Immunization since 2002. We aimed to survey the seroprevalence of and immunity to hepatitis B virus (HBV) in children born from 2002 to 2014 in Jiangsu, China.MethodsTotally 3442 children (M:F = 2072:1370) at the age of 7 months to 12 years (5.5 ± 3.6), from five cities and rural areas across Jiangsu province, were enrolled. Blood samples were measured for HBV markers by ELISA and quantitative microparticle enzyme immunoassay. HBV DNA was tested by real-time PCR and S region was amplified by nested PCR.ResultsTwelve (0.35%) children were positive for hepatitis B surface antigen (HBsAg) and 34 (0.99%) were HBsAg negative and positive for antibody against hepatitis B core antigen (anti-HBc). Totally 2542 (73.85%) children had anti-HBs levels ⩾10 mIU/ml and 535 (15.54%) with 2–9.9 mIU/ml. All 12 HBsAg-positive children had detectable HBV DNA with a mean level of 6.1 ± 1.7 log IU/ml (3.3–8.1 log IU/ml); 8 were genotype C and 4 were genotype B. No mutation was detected in the a determinant of HBsAg. HBV DNA was not detected in all the 34 children with positive anti-HBc and negative HBsAg.ConclusionHBsAg prevalence among children in Jiangsu born after the introduction of universal vaccination against hepatitis B has significantly decreased. No mutation of S gene is associated with vaccine failure in the cohort of children.     

2003～2005年永州市青少年学生乙肝感染动态分析

目的分析近3年来乙肝病毒在不同类群青少年学生中的感染状况及动态变化。方法以酶联免疫吸附试验（ELISA）法，检测2003-2005年永州市参加中考、高考各类青少年学生共7605名的乙肝表面抗原（船sAg）及“乙肝两对半”指标，参考其他因素和其他城市情况，逐年分组进行分析。结果3年船sAg平均阳性率为11．65％，2003～2005年每年的船sAg阳性率分别为10．20％、11．75％、13．36％，男女生阳性率分别为12．10％、10．28％，城乡学生阳性率分别为10、92％、13．16％，住校生、走读生阳性率分别为12．83％、11．15％，初、高中毕业生阳性率分别为10．88％、12．01％。886例HgsAg阳性学生中乙肝e抗原阳性者占61．52％。结论我市中青少年学生的乙肝感染率不但偏高，而且有不断增高趋势，其中传染性强的人群较大。农村学生、男生、住校生为乙肝重点感染人群，需给予特别关注。