重组乙型肝炎病毒变异s基因疫苗诱导小鼠产生特异性体液免疫应答

目的：构建乙型肝炎病毒(HBV)变异s基因真核表达载体，检测其诱导小鼠产生特异性体液免疫应答。方法：利用限制性内切酶定位克隆构建s基因nt587 G→A的真核表达载体pcMV-S2．S 145R(PR).用其转染人肝癌细胞系Hep G2后，用EIA、EILISA及免疫细胞化学法，观察其抗原性。以重组变异型s基因真核表达载体(PR)和载体pcDNA3．0分别免疫C57BL／6小鼠各5只。每只小鼠各肌肉注射纯化质粒100μg.用ELISA法检测血清抗-HBs及抗-HBs2抗体的效价。结果：体外实验证实，变异型HBs矩可与抗-HBs结合；PR免疫小鼠可诱导其产生抗-HBs抗体及抗．HBs2抗体，但抗-HBs2抗体的出现早于抗-HBs抗体约1～2wk。结论：HBV变异s基因(nt587G→A)的真核表达载体的表达产物具有良好的抗原性，能够诱导C57BL／6小鼠产生体液免疫应答。     

新生儿乙型肝炎疫苗免疫后远期效果的研究

对湘潭市市区０－１０岁儿童乙型肝炎（乙肝）疫苗免疫后的效果进行了系统监测。６次共调查儿时１２２名，各次调查的乙型肝炎病毒表面抗原（ＨＢｓＡｇ）携带率在０．００１－１．８３％之间，平均ＨＢｓＡｇ携带率１．１２％，明显低于免疫前现一人群的１６．１７％，保护率为９２．４６％。初次免疫后抗－ＨＢｓ的阳经及几何平均滴度（ＧＭＴ）高峰值均出现在６－１２个月，此后随时间的推移而逐渐下降，但７－１０岁儿童中仍有５     

乙型肝炎疫苗加强接种2年后的免疫应答

<正> 我们曾报道对乙型肝炎疫苗(下简称乙肝疫苗)初免3年抗-HBs呈低水平(S/N≤5)和转阴的儿童,加强注射1剂10μg或30μg乙肝疫苗,取得快而好的回忆反应,本文报道加强免疫2年后受检查者抗-HBs水平及保护力的观察结果。 材料和方法 观察对象的选择和分组均见前报,新生儿(3岁)组随机分成二组,分别加强接种1剂10μg或30μg乙肝疫苗,儿童(4～8岁)组     

CCR5基因多态性与乙型肝炎病毒感染相关性的研究进展

趋化性细胞因子和趋化性细胞因子受体在抗病毒免疫中发挥了重要作用。乙型病毒性肝炎是由其表达的抗原系统及其抗体所介导的特异性免疫反应和非特异性的以细胞因子为主的炎症介质导致的肝细胞损伤。CCR5作为趋化性细胞因子受体在乙型病毒性肝炎的发生发展中起重要作用。本文主要讲述CCR5基因的结构和功能,及在编码区和启动子区的多态性与HBV感染的相关性研究。     

过敏反应与细胞因子及其基因多态性

过敏反应的发病机制非常复杂 ,受多种因素影响 ,其中Th1 /Th2平衡失调 ,以及体内细胞因子都在过敏反应中发挥重要作用。在细胞因子中白细胞介素类、趋化性细胞因子、干扰素、集落刺激因子和肿瘤坏死因子等几大类约 3 0余种细胞因子 (IL 4、IL 5、IL 1 0、IL 1 2、IL 1 8、IFN γ、TNF α、TGF β、RANTES、Eotaxin、MCP 1等 )在介导过敏反应的细胞免疫、体液免疫和过敏反应的调节中起重要作用。这些细胞因子的受体 (IL 4R、IL 1 3R、CCR3、CCR5等 )也与过敏反应有关。　　过敏反应有明显的个体差异和遗传倾向 ,一般认为过…     

纤维介素基因启动子的基因多态性与重型乙型肝炎关系的研究

近年小鼠暴发性肝炎和人重型肝炎的研究表明 ,纤维介素 (fgl2 )的高表达是重型乙型肝炎肝细胞坏死的重要分子机制。我们运用单链构象多态性 (singlestrainconstructionpolymorphism ,SSCP)分析方法对重症乙型肝炎患者、乙型肝炎病毒携带者以及健康人hfgl2基因启动子的基因多态性进行了研究。4 0例标本均收集于华中科技大学同济医学院附属同济医院感染科 ,其中重型乙型肝炎患者 2 1例 (男性 14例 ,女性 7例 ,平均年龄 37.5岁 ) ,乙型肝炎病毒携带者 15例 (男性 10例 ,女性 5例 ,平均年龄 33.2岁 ) ,健康人 4例 (男性 3例 ,女性 1例 ,平均年…     

全身性感染与细胞因子基因多态性的相关性

机体在遭受细菌或毒素损害时,炎症细胞被激活,细胞因子过量释放,产生持续的全身性炎症反应,介导广泛的组织损伤,导致感染性休克和多器官功能衰竭.细胞因子的基因多态性与机体受到刺激后细胞因子的分泌水平相关,与全身性感染的易感性、严重程度及预后相关.探明细胞因子基因多态性与全身性感染的关系,将为全身性感染的个体化免疫基因调控治疗奠定基础.     

乙肝核酸疫苗NV—HB／s接种后小鼠体液免疫应答的初步研究

乙肝核酸疫苗ＮＶ┐ＨＢ／ｓ接种后小鼠体液免疫应答的初步研究赵连三秦山李水仙唐红刘丽周思亮雷秉钧将乙肝病毒表面抗原（ＨＢｓＡｇ）主蛋白的编码基因Ｓ插入真核细胞表达质粒ｐＲｃ／ＣＭＶ／ＡＴＧ中的ＣＭＶ启动子下游，获得的重组质粒（ＮＶ－ＨＢ／ｓ）转染真核细...     

过敏性紫癜细胞因子基因多态性的研究进展

过敏性紫癜（HSP）是儿童期较为常见的小血管性血管炎,其病因及发病机制尚不明确,近年来有研究表明某些细胞因子的基因多态性与HSP的发病机制、易感性、病理进展及预后等密切相关,如自细胞介素基因、细胞粘附分子基因、血管内皮生长因子基因、转化生长因子基因、肿瘤坏死因子基因等。因此对与HSP相关的细胞因子基因多态性进行综合分析将有着重要意义,为今后临床上从基因水平认识HSP疾病的本质及开发新的研究和新的治疗方案提供了重要的依据。     

GM—CSF促进乙肝病毒基因疫苗诱导的抗体产生

将构建的编码乙肝病毒PreS2+S蛋白的真核表达质粒注射于C57BL/6小鼠胫前肌内,3d后,在同一部位注射rhGM-CSF。基因疫苗接种2周后,血清中可测到抗-HBs,两月后,抗体水平达到高峰,并保持高水平至少2月以上;肌内注射GM-CSF可提高血清抗-HBs水平,提示GM-CSF可作为基因疫苗的佐剂。     

乙肝疫苗免疫耐受性相关基因研究进展

乙肝疫苗接种可刺激机体免疫系统产生保护性抗体,但部分人群接种后并没有有效的抗体产生.发生乙肝疫苗低或无应答的免疫耐受因素众多,机体的遗传因素是一个重要原因.该文就接种乙肝疫苗后所产生的免疫耐受现象与机体遗传相关基因的研究进展进行综述.     

Qualitative analysis of the humoral immune response to the "a" determinant of HBs antigen after inoculation with plasma-derived or recombinant vaccine

Competitive inhibition assays using monoclonal antibodies reacting with defined sequence of the "a" determinant of hepatitis B surface antigen (HBsAg) indicate that the humoral response to plasma-derived and recombinant HBsAg vaccine is similar in quantitative and qualitative terms. Both vaccines evoke an antibody response to the RFHBs 1 epitope, which is known in animal experiments to be protective.     

CpG ODN联合乙型肝炎疫苗与乙型肝炎疫苗免疫效果对比研究

目的探讨CpG ODN乙型肝炎疫苗与市售乙型肝炎疫苗对Balb/c小鼠的免疫作用效果。方法将乙肝疫苗、乙肝疫苗+100μgCpG ODN分别肌肉注射到4～6周龄,16～18g Balb/c小鼠体内,于第1次免疫后28d以同样剂量加强免疫1次。分别于第1次免疫后28、42、63 d收集小鼠血清,用ELISA方法检测抗HBs IgG抗体。结果实验组产生的抗HBs IgG与对照组HBsIgG相比P<0.05,具有显著性差异。28、42、63 d抗HBs IgG分别是对照疫苗的114倍、4.73倍、5.41倍以上。结论 CpG ODN能够显著增强小鼠对乙肝疫苗的免疫应答,在较短时间内显著的提高乙肝抗体的产生水平,免疫效果是乙肝疫苗的5倍以上。     

Association between tumour necrosis factor gene polymorphisms and the clinical types of patients with chronic hepatitis B virus infection

A PCR restriction fragment length polymorphism assay was used to analyse single-nucleotide polymorphisms in the tumour necrosis factor (TNF)-alpha and TNF-beta genes of 56 patients with chronic severe hepatitis B virus (HBV) infection, 71 patients who either had chronic mild HBV infection or who were asymptomatic carriers, and 90 healthy controls. The serum TNF-alpha concentrations in patients with chronic severe HBV infection were compared to those of 30 healthy controls by radioimmunoassay. The frequencies of the TNF1/2 genotype and the TNF2 allele were greater in patients with chronic severe HBV infection than in healthy controls (25% vs. 11.1%, p 0.015; 12.5% vs. 5.6%, p 0.036, respectively) and patients with chronic mild HBV infection and asymptomatic carriers (25% vs. 8.8%, p 0.011; 12.5% vs. 4.2%, p 0.015, respectively). Heterozygotes carrying the TNF2 allele had higher levels of serum TNF-alpha than homozygotes for the wild-type allele among all patients with chronic severe HBV infection (p <0.01). The genotype distribution and allele frequency of TNF-beta were similar for patients with chronic severe HBV infection and healthy controls, but the frequency of the TNF-beta*2/2 genotype in patients with chronic mild HBV infection and asymptomatic controls was lower than for healthy controls (9.9% vs. 22.4%, p 0.043) or patients with chronic severe HBV infection (9.9% vs. 26.8%, p 0.043), although this was not significant after correction for multiple testing. It was concluded that TNF-alpha gene polymorphisms may play an important role as a host factor in the progression of HBV infection.     

Association of cytokine genetic polymorphisms with the humoral immune response to recombinant vaccine against HBV in infants

The prevention of hepatitis B by vaccination is one the most efficient tools to avoid the transmission of the virus, although a considerable variability to the anti‐HBsAg antibody response has been described. Recently, polymorphisms of cytokine regulating genes have been described which seem to influence the immune response to various antigens. This article's objective was to evaluate the influence of cytokine genetic polymorphisms onto the humoral immune response to hepatitis B vaccine in infants. Vaccinated children were classified according to the level of anti‐HBsAg antibody titles. The genotyping for TNF (?308), TGFB1 (+869, +915), IL‐10 (?1082, ?819, ?592), IL‐6 (?174), and IFNG (+874) was accomplished by the PCR‐SSP technique. The TNF (?308) allele A presented a lower but not statistically significant frequency at 5% level in high responder patients (3.7% vs. 12.3%, P = 0.0919). The same was seen for the TNF (?308) genotype GA (7.4% vs. 24.5%, P = 0.0757). Further studies in other populations and evaluation of a greater number of individuals may contribute for a better understanding of the cytokine gene polymorphism influence in general and TNF polymorphism more specifically in the humoral immune response to the HBsAg vaccination in newborn children. J. Med. Virol. 82:929–933, 2010. © 2010 Wiley‐Liss, Inc.     

Association between polymorphisms of the cytokine and cytokine receptor genes and immune response to hepatitis B vaccination in a Chinese Han population

The immune response to hepatitis B vaccination varies among individuals. It has been reported that polymorphisms in cytokine and cytokine receptor genes are associated with these individual differences. The aim of the current study was to investigate the association between polymorphisms of the Th1/Th2 cytokine and cytokine receptor genes and the response to hepatitis B vaccination in a Chinese Han population. A total of 10 single nucleotide polymorphisms distributed in 6 genes (TNFRSF1A, IL12A, IL12B, IFNG, IL4, and IL10) were genotyped in 214 high‐responders [hepatitis B surface antibody (anti‐HBs) ≥1,000 mIU/ml] and 107 low‐responders (anti‐HBs: 10–99 mIU/ml). The minor CTCTAA allele of rs17860508 in the IL12B gene was associated with a low response to hepatitis B vaccination (P = 0.039, odds ratio = 1.41, 95% confidence interval = 1.00–1.99). In addition, a significant gene–gene interaction was found: the frequency of the combined genotypes IL12A rs2243115 TT and IL12B rs17860508 CTCTAA/CTCTAA was significantly higher in the low‐response group than in the high‐response group (P = 0.008, odds ratio = 2.19, 95% confidence interval = 1.23–3.93). These findings suggest that polymorphisms in the IL12A and IL12B genes might play an important role jointly in determining the response to hepatitis B vaccination. J. Med. Virol. 84:26–33, 2011. © 2011 Wiley Periodicals, Inc.     

Association of apolipoprotein E with the progression of hepatitis B virus-related liver disease

Hepatitis B virus (HBV) infection increases the risk of liver decompensation, cirrhosis and hepatocellular carcinoma (HCC). Apolipoprotein E (ApoE), one of the major cholesterol carriers, plays important role in the metabolism of lipoprotein and the regulation of immune response. The present study was aimed to explore whether the genetic variation in ApoE gene affected disease progression in HBV infected individuals. We collected sera samples from healthy volunteers (n=40), inactive HBV carriers (n=30), and patients with acute hepatitis (n=60), severe hepatitis (n=12), HBV-related liver cirrhosis (n=58) or primary HCC (n=39). We found that ApoE and interlukin-6 (IL-6) was progressively increased, while IL-2 was gradually decreased with the increasing grade of disease severity. Furthermore, high ApoE levels in HBV infected individuals were correlated with increased IL-6 and decreased IL-2 levels, indicating immune abnormalities in these patients. The frequency of E3/3 genotype was progressively increased from carriers group, hepatitis group to progressive group (cirrhosis and HCC). The serum levels of low-density lipoprotein cholesterol (LDL-C) differed among ApoE phenotypes, with E3/4, E4/4> E3/3>E2/3. Our study suggested that ApoE may have a role in the pathogenesis and progression of HBV-related liver disease and indicated the possible underlying mechanisms.     

IL-28B基因多态性与丙型肝炎相关性研究进展

白细胞介素-28 B(Interleukin-28B,IL-28 B)属于Ⅲ型干扰素家族,是一种新型的白细胞介素.它通过活化JAK- STAT信号通路,调节干扰素刺激基因转录,发挥抗病毒等生物学效应.新近研究发现,IL-28B基因多态性与丙型病毒性肝炎的发病、抗病毒治疗应答和疾病转归等密切相关.因此,通过对二者相关性的研究,可能有助于临床医师更合理的选择丙型肝炎的治疗措施、开展个性化治疗、及早判断患者的预后.     

Myxovirus resistance,osteopontin and suppressor of cytokine signaling 3 polymorphisms predict hepatitis C virus therapy response in an admixed patient population: comparison with IL28B

OBJECTIVES:

Suppressor of cytokine signaling 3, myxovirus resistance protein and osteopontin gene polymorphisms may influence the therapeutic response in patients with chronic hepatitis C, and an association with IL28 might increase the power to predict sustained virologic response. Our aims were to evaluate the association between myxovirus resistance protein, osteopontin and suppressor of cytokine signaling 3 gene polymorphisms in combination with IL28B and to assess the therapy response in hepatitis C patients treated with pegylated-interferon plus ribavirin.

METHOD:

Myxovirus resistance protein, osteopontin, suppressor of cytokine signaling 3 and IL28B polymorphisms were analyzed by PCR-restriction fragment length polymorphism, direct sequencing and real-time PCR. Ancestry was determined using genetic markers.

RESULTS:

We analyzed 181 individuals, including 52 who were sustained virologic responders. The protective genotype frequencies among the sustained virologic response group were as follows: the G/G suppressor of cytokine signaling 3 (rs4969170) (62.2%); T/T osteopontin (rs2853744) (60%); T/T osteopontin (rs11730582) (64.3%); and the G/T myxovirus resistance protein (rs2071430) genotype (54%). The patients who had ≥3 of the protective genotypes from the myxovirus resistance protein, the suppressor of cytokine signaling 3 and osteopontin had a greater than 90% probability of achieving a sustained response (p<0.0001). The C/C IL28B genotype was present in 58.8% of the subjects in this group. The sustained virological response rates increased to 85.7% and 91.7% by analyzing C/C IL28B with the T/T osteopontin genotype at rs11730582 and the G/G suppressor of cytokine signaling 3 genotype, respectively. Genetic ancestry analysis revealed an admixed population.

CONCLUSION:

Hepatitis C genotype 1 patients who were responders to interferon-based therapy had a high frequency of multiple protective polymorphisms in the myxovirus resistance protein, osteopontin and suppressor of cytokine signaling 3 genes. The combined analysis of the suppressor of cytokine signaling 3 and IL28B genotypes more effectively predicted sustained virologic response than IL28B analysis alone.     

HLA-DRB1等位基因与吉林地区汉族乙肝疫苗免疫应答的关联研究

目的:探讨HLA-DRB1等位基因与吉林地区汉族乙肝疫苗免疫应答的关联性。方法:采用聚合酶链反应-序列特异性引物(PCR-SSP)分型技术,对84例乙肝疫苗无或低应答者HLA-DRB1等位基因进行检测,与78例乙肝疫苗中或强应答者人群进行对照。结果:①HLA-DRB1*14等位基因频率在无或低应答组为23.8%,中或强应答组为5.13%,两组之间比较有统计学差异(P<0.05);②HLA-DRB1*12、HLA-DRB1*15等位基因频率分别在无或低应答组为4.76%和7.14%,在中或强应答组为23.1%和24.4%,两组之间比较也存在统计学差异(P<0.05)。③等位基因HLA-DRB1*07(2.38%和5.12%),HLA-DRB1*08(9.52%和8.97%),HLA-DRB1*09(7.14%和10.3%),HLA-DRB1*11(7.14%和7.69%),HLA-DRB1*13(4.76%和6.41%)及HLA-DRB1*16(4.76%和5.13%),在无或低应答组和在中或强应答组之间基因频率分布没有统计学差异(P>0.05)。结论:吉林地区汉族乙肝疫苗接种后,①HLA-DRB1*14等位基因可能与无或低应答相关;②HLA-DRB1*12、15等位基因可能与中或强应答相关;③未检测到HLA-DRB1*07、08、09、11、13、16等位基因与免疫应答水平之间有明显的相关性。