多表位病毒疫苗的研究进展

多表位病毒疫苗是目前病毒疫苗研制的特点,特别是对目前病毒疫苗研制的热点,特别是对于易变异或者本身有癌基因的病毒。本文就近年来这方面的研究进展作一综述。     

表位疫苗研究进展

表位疫苗是用抗原表位制备的疫苗,是近年来新兴的一种疫苗研制技术,也是今后最具开发前景的疫苗技术之一,在肿瘤、病毒等疾病的防治中有着自身独特的优势.详细阐述了T表位和B表位的筛选和鉴定方法、表位疫苗的载体研究及表位疫苗在肿瘤、病毒和微生物感染中的应用等,对表位疫苗的最新研究进展进行了综述.     

确定病毒B细胞抗原表位的策略

抗原表位是诱生免疫反应的抗原最基本单位,根据其功能的不同,可大致分为Ｂ细胞、Ｔｈ细胞及Ｔｃ细胞抗原表位。如何确立这些抗原表位是近年分子免疫学研究的热点。本文综述了有关确立病毒Ｂ细胞抗原表位的方法及策略,并评价了这些方法的优缺点及适用范围。     

表位疫苗及相关技术研究进展

表位疫苗是用抗原表位制备的疫苗,是近年来新兴的一种疫苗研制技术,也是今后最具开发前景的疫苗技术之一,在肿瘤、病毒等疾病的防治中有着自身独特的优势。本文详细阐述了T表位和B表位的筛选和鉴定方法、表位疫苗的载体研究及表位疫苗在肿瘤、病毒和微生物感染中的应用等,对表位疫苗的最新研究进展进行了综述。     

T细胞抗原表位预测的研究方法进展

确定T细胞所识别抗原分子上的短肽序列对T细胞表位进行定位,对于研究特异性免疫应答有着重要意义。综述了近年来实验确定和理论预测T细胞蛋白质抗原袁位的常用方法,以及T细胞抗原表位分析的研究方法。     

B 细胞抗原表位的研究进展及其应用

抗表位是抗分子中决定抗特异性的特殊化学基团,其对研究特异性免疫应答有着重要意义.简要综述了蛋白质抗表位的种类及特性,回顾了近几年来理论预测和实验确定 B 细胞抗表位的常用方法及 B 细胞抗表位分析的研究方法,以及抗表位在流感流行预测及疫苗安全性方面的应用.     

B细胞表位预测方法研究进展

B细胞抗原表位预测方法的研究对基础免疫学的研究及实际应用有着重要的意义。本文归纳了理论预测B细胞表位的常用方法,并对目前预测B细胞表位方法存在的问题进行了分析。     

HCV复合多表位抗原基因表达及免疫原性的研究

分类号Ｑ７８１文献标识码Ａ文章编号０００１６２０９（１９９９）０３０２６８７１丙型肝炎是常见的传染病之一,易于慢性化及发展为肝硬化,且与肝癌的发生关系密切,目前尚无理想的防治手段。国内外对丙型肝炎疫苗的研究仍处于克隆表达ＨＣＶ部分基因产物或合成…     

蛋白质抗原表位研究进展

本文综述了蛋白质抗原表位的种类及特性,回顾了近几年来实验确定和理论预测B细胞蛋白质抗原表位的常用方法,介绍了表位作图和表位疫苗的研究现状。     

丙型肝炎病毒NS3蛋白辅助T细胞表位研究

对２例ＨＣＶ持续性感染者２个时间点ＮＳ３区Ｃ末端克隆测序,研究ＨＣＶ ＮＳ３蛋白一个辅助Ｔ细胞表位（氨基酸位１２４８－１２６１）在ＨＣＶ感染者体内的保守性;人工合成该表位进行淋巴细胞增殖试验和抑制试验,观察该表位引起免疫应答的水平和特点;通过“刺激－休息－刺激”多轮循环建立该表位特异性的Ｔ细胞系,并用流式细胞仪鉴定表型。结果发现该表位在２个病人２个时间点均未变化;观察的２例ＨＣＶ持续性感染者和１例     

B Cell Epitopes within VP1 of Type O Foot-and-mouth Disease Virus for Detection of Viral Antibodies

In this study,the coding region of type O FMDV capsid protein VP1 and a series of codon optimized DNA sequences coding for VP1 amino acid residues 141-160(epitope1),tandem repeat 200-213(epitope2(+2)) and the combination of two epitopes(epitope1-2)was genetically cloned into the prokaryotic expression vector pPROExHTb and pGEX4T-1,respectively.VP1 and the fused epitopes GST-E1,GST-E2(+2)and GST-E1-2 were successfully solubly expressed in the cytoplasm of Escherichia coli and Western blot analysis demonstrat...     

猪口蹄疫病毒多抗原表位重组腺病毒的构建与鉴定

本研究设计构建了含有猪O型口蹄疫病毒VP1（21—60）-（141-160）-（200—213）位氨基酸的基因的重组腺病毒质粒pAd-VP,经PacI酶切后转染HEK-293A细胞,3次噬斑纯化获得了重组腺病毒rAd—VP。该重组腺病毒于HEK-293A细胞连续传代至20代效价稳定,TCID50为10^-10／mL。RT—PCR检测证明目的基因在mRNA水平上可有效表达;应用O型口蹄疫病毒标准阳性血清进行间接荧光抗体试验,在rAdVP感染的HEK-293A细胞的胞质可见清晰荧光。证明该重组腺病毒对VP1（21-60）-（141—160）-（200—213）位氨基酸的基因进行了成功的表达,从而为FMDV多抗原表位腺病毒活载体疫苗的研究奠定了基础。     

Develope Monoclonal Antibody against Foot-and-mouth Disease Virus A Type

In order to develop an anti-FMDV A Type monoclonal antibody (mAb),BABL/c mice were immunized with FMDV A type.Monoclonal antibodies (mAbs) 7B11 and 8H4 against Foot-and-mouth disease virus (FMDV) serotype A were produced by fusing SP2/0 myeloma cells with splenocyte from the mouse immunized with A/AV88.The microneutralization titer of the mAbs 7B11 and 8H4 were 1024 and 512,respectively.Both mAbs contain kappa light chains,the mAbs were IgG1.In order to define the mAbs binding epitopes,the reactivity of the...     

口蹄疫病毒VP1基因转化马铃薯及其块茎专一性表达

报道了FMDV VP1基因与马铃薯块茎专一性表达class Ipatatin基因5′区融合,经农杆菌介导导入马铃薯植株,PCR、RT-PCR证实了其整合及转录表达。ELISA结果进一步表明,VP1在转基因马铃薯块茎中具有免疫活性。为探讨在马铃薯块茎中高表达VP1蛋白及进一步开发其作为FMDV口服疫苗生物反应器奠定基础。     

Asia1型口蹄疫病毒第V群猪源和牛源毒株全基因组比较分析

为了查明Asia1型FMDV第Ⅴ群猪源和牛源毒株的序列差异, 采用RT-PCR方法, 对Asia1型FMDV第Ⅴ群猪源分离毒株Asia1/HN/06的基因组全序列进行了扩增和测序, 并与第Ⅴ群牛源和猪源参考毒株基因组进行比较分析。结果表明, Asia1/HN/06毒株全基因组序列长约8236 nt [含38个A的poly(A)尾], 其中5'NCR长1116 nt, 前导蛋白(L)编码区长603 nt, 结构蛋白与非结构蛋白编码区的核苷酸序列为6990 nt, 3'NCR长93 nt, 3￠端是至少含有38个A的poly(A)尾巴。猪源毒株和牛源毒株的全基因比较分析表明, 属于第Ⅴ群, 全基因编码区核苷酸和氨基酸的同源性均为98.0%, 主要差别是猪源毒株Asia1/HN/06在细胞受体结合位点变为RDD和155位置的N变为S或D, 该群毒株3A更具有猪源毒株特征, 有4个特异性氨基酸变异。明确了Asia1型FMDV第Ⅴ群猪源和牛源毒株的序列差异, 为进一步利用反向遗传技术研究猪源和牛源毒株差异位点或基因在病毒表型变异中的作用奠定基础。     

口蹄疫病毒株AF72 VP3的结构模拟与分析

以口蹄疫病毒株AF72 RNA为模板,反转录并扩增目的基因,PCR纯化产物与pGEM TEasy载体连接并转化JM109菌株,用凝胶电泳、PCR和Spe I/Sph I双酶切法鉴定为阳性的重组质粒进行测序.比对测序结果确定AF72 VP3的核苷酸序列,利用同源建模的方法建立AF72 VP3结构蛋白的3D结构,在此基础上,综合亲水性、可塑性、抗原指数以及表面可能性等参数预测AF72 VP3结构蛋白的B细胞抗原表位.分析表明,口蹄疫病毒VP1、VP2、VP3和VP4在核苷酸水平上的变异率是无差异的(P＞0.05);而它们在氨基酸水平上的变异率差异显著(P＜0.05).该毒株与20株源于GenBank中的VP3氨基酸序列比对发现其保守区主要位于第1～24、24～35、36～42、45～56、65～122、124～172、177～210、211～219位.AF72 VP3结构蛋白三维空间结构可分为A、B和C 3个结构区域,蛋白呈现较规则的空间构象,其中18～23、30～44、60～75、113～124、130～142、193～220氨基酸区段是AF72VP3结构蛋白可能的B细胞抗原表位区域,该结果将为进一步的FMDV多表位疫苗研究提供更有价值的参考信息.     

Application of VP1 Protein to Develop Monoclonal Antibody against Foot-and-mouth Disease Virus Asial Type

In order to develop an anti-FMDV Asial type monoclonal antibody (mAb), BABL/c mice were immunized with recombinant FMDV VP1 protein. Three mAbs, 1B8, 5E1 and 5E2, were then further optimized. The result indicated that prepared anti-FMDV Asial mAbs had no cross-reactivity with Swine vesicular disease (SVD) and FMDV O, A and C type antigen. Their titers in abdomen liquor were l:5×106, l:2×106 and l:5×l06, respectively. 1B8 was found to be of IgGi subtype, 5E1 and 5E2 belonged to IgG2b subtype. In this study, the prepared mAbs are specific for detecting FMDV type Asial, and is potentially useful for pen-side diagnosis.     

Amplification and Characterization of Bull Semen Infected Naturally with Foot-and-mouth Disease Virus Type Asial by RT-PCR

To investigate the security of semen biologically, 15 bull semen samples were collected (of which 5 exhibited clinical signs of Foot-and-mouth disease) and identified by RT-PCR and virus isolation. The results indicated that the semen of the infected bulls were contaminated by Foot-and-mouth disease virus (FMDV), but FMDV was not detected in semen samples from those bulls not showing clinical signs of Foot-and-mouth disease (FMD). This is the first report of the presence of FMDV in bull semen due to natural infection in China. The analysis of the partial sequence of the VP1 gene showed that the virus strain isolated from semen has 97.9% identity with the virus isolated from vesicular liquid of infected bulls showing typical signs of FMD and belonged to the same gene sub-group.     

The Solution Structure of the Immunodominant and Cell Receptor Binding Regions of Foot-and-mouth Disease Virus Serotype A,Variant A

The solution structure of a 20 amino acid long peptide corresponding to the region 141–160 of the envelope protein Vp1 from foot-and-mouth disease virus (FMDV) serotype A, variant A, has been determined by a combination of NMR experiments and computer calculations. The peptide contains both the immunodominant epitope as well as the sequence (RGD) used by the virus to bind the cell receptor in the initial stages of infection. These two sites have been shown to partially overlap. One hundred and thirty-five NMR distance constraints were used to obtain a set of 11 structures by distance geometry, minimization and molecular dynamics simulations. These structures were divided into two homogeneous families based upon backbone superimposition. The first and most populated family was characterized by a backbone RMS of 1.5±0.4 Å, the second by a backbone RMS of 0.8±0.2 Å. The two families had similar structural features and differed mainly in the backbone angles of G149. In the larger of the two families these angles favoured the formation of a loop comprising residues 147 to 152 and stabilized by a H-bond between the NH of D147 and the CO of A152. In the second family, where this bond was absent, the peptide adopted in this region the shape of an irregular helix. The C-terminal half of the peptide (152–159) was similar in both families and largely helical. Similar structural features were also found within the VRGDS sequence (144–148) which was assigned to a β-turn type IV. The features of the two families of structures were found to be different from those of the recently published X-ray structure of the antigenic loop of a chemically modified form of FMDV. Proposals accounting for these differences are provided which take into account the dual activity of the 141–160 sequence (i.e. antibody binding and cell invasion through receptor binding).     

整联蛋白与口蹄疫病毒感染

整联蛋白是一类分布广泛的细胞表面受体家族,它不仅在细胞的生长、移行、增殖和分化等许多方面发挥重要生物学功能,而且在多种病理过程中发挥重要作用。许多病毒都能利用整联蛋白分子作为病毒受体或共受体进入宿主细胞。本文主要就整联蛋白的特征及其在口蹄疫病毒感染宿主细胞过程中的作用机制进行综述。