细菌菌蜕的研究进展

细菌菌蜕是革兰阴性细菌被噬菌体PhiX174的裂解基因E裂解后形成的完整细菌空壳.这种基因灭活过程其内外膜结构保持良好,不会引起细菌表面结构的任何物理化学变化,可作为递送系统和疫苗使用,在疾病的预防和治疗方面具有广阔的应用前景.     

表面等离激元共振成像技术在识别和确认诱导多能干细胞生物标记中的应用(英文)

诱导多能干细胞(i PSCs)是再生医学领域,尤其是细胞治疗和药物筛选研究中非常有吸引力的细胞来源.然而,再生医学需要对i PSCs进行快速、精准鉴别.在本研究中,我们开发了针对多能性生物标记物的抗体阵列,分别以胚胎干细胞(ESCs)和鼠成纤维细胞(MEF)裂解液作为阳性和阴性对照,应用快速且无需标记的表面等离激元共振成像(SPRi)技术对i PSCs裂解液中的多种抗原进行检测.每一种抗体都与i PSC裂解液中相应的抗原显示出特异性识别.这一结果表明,SPRi技术适合于检测i PSCs裂解液中的多种抗原,并具有通用性,利用SPRi技术可以用于对干细胞裂解液中生物标志物的高通量鉴别分析.     

TritonX-100裂解流感病毒的研究

 在流感病毒高浓度条件下,寻找TritonX-100裂解流感病毒的适宜条件.在不同条件下用TritonX-100裂解WHO推荐用2007～2008年度流感病毒疫苗毒株制备的3个亚型的流感病毒,用电镜观察裂解程度,通过蔗糖密度梯度离心纯化抗原后免疫动物,以检测疫苗的免疫效果.结果表明当流感病毒的血凝效价是1:16382,裂解时间为24h时,TritonX-100在质量分数为1%可以完全地裂解B型,质量分数达到2%才可以完全裂解流感病毒H1N1和H3N2,这说明TritonX-100对A,B亚型毒株的裂解效果不同.由于每年都要更换毒株工人生产新的流感疫苗,因此应该对使用TritonX-100的裂解不同亚型的流感病毒条件进行研究,以保证裂解疫苗的质量.     

用合成的碳水化合物制造疫苗

科学家合成了一种安全、有效的基于碳水化合物的疫苗 ,这个疫苗是针对一种引起髓膜炎的细菌的。这一合成碳水化合物疫苗的成功显示 ,有了合成碳水化合物的生产 ,人们也许能较廉价和容易地制造针对其它细菌疾病的疫苗。文章作者报告说 ,这个疫苗和靠从细菌采集碳水化合物制造的传统疫苗效果一样。专家们评价这是第一个大规模生产的、并经临床评估的合成碳水化合物疫苗。他们描述了用一个来自 B型流感嗜血杆菌的合成碳水化合物抗原批量成产和评估疫苗的过程。用合成的碳水化合物制造疫苗     

新型免疫疫苗——DNA疫苗

DNA疫苗是近几年发展起来的一种新型疫苗 .将抗原编码基因插入带有强启动子的质粒载体,然后用物理方法将重组质粒导入体内细胞,抗原编码基因即在细胞内合成抗原蛋白,诱发机体产生保护性免疫反应 .DNA疫苗是独特而有效的接种方法,实验证明它可以用来抵抗病毒感染和医治肿瘤等,有广泛的应用前景 .     

多杀性巴氏杆菌幽灵的研究

采用基因工程方法,将PhiX174噬菌体裂解基因E和温敏控制表达系统与质粒pPBA1100基因杂交,构建了重组子pPBA1100-E.将重组子转化到多杀性巴氏杆菌中,通过温度诱导使裂解基因表达.用限制性内切酶检验重组子.扫描电镜观察多杀性巴氏杆菌细菌幽灵和菌落形成单位评价遗传灭活率.结果表明:重组质粒经限制性内切酶酶切鉴定,琼脂糖电泳呈现三条谱带,相对分子质量与理论值一致.扫描电镜观察显示,重组子在多杀性巴氏杆菌中成功表达,获得了细菌幽灵.菌落形成单位检验结果显示,多杀性巴氏杆菌遗传灭活率达到99%.     

不同动物血清补体对大肠杆菌的杀灭效果

探索不同动物血清补体对大肠杆菌的杀灭效果可以为开发利用补体作为新型疫苗佐剂提供理论依据。将培养活化的O78大肠杆菌用甲醛灭活,以蜂胶为佐剂制备大肠杆菌颗粒性疫苗,用其免疫11日龄海蓝褐蛋用公雏,制备大肠杆菌抗血清;将大肠杆菌抗血清灭活补体,与已知数量的大肠杆菌反应,形成抗原抗体复合物,然后加入不同动物的新鲜血清,细菌计数,观察不同动物血清补体对大肠杆菌的清除能力;微量板法测定不同动物血清补体总活性。结果表明,抗体与细菌抗原结合后,对细菌的杀灭作用较小,动物体内的补体,是细菌性疫苗发挥免疫效果的关键物质;哺乳动物血清中的补体对细菌的清除能力高于鸡,在哺乳动物中,豚鼠的血清补体对大肠杆菌的清除能力最高。     

猪型布鲁氏菌Omp31抗原表位基因的性质分析

布鲁氏菌病是一种由革兰氏阴性小杆菌:布鲁氏菌(Brucella)引起的,主要在动物中以流产和发热为表现特征的疾病.以猪型布鲁氏菌病Omp31抗原表位基因作为研究对象,对基因的种间同源性、亚型分类、结构性质、抗原表位性质等信息进行了分析,并且作出了相关预测.随着预测方法的不断完善,将会对布鲁氏菌病其它抗原表位基因的研究提供帮助,从而为该疾病的防疫、治疗提供理论依据,同时也可以促进相关疫苗的研制.     

细菌内毒素对基因免疫小鼠存活率和产仔率的影响

作为第三代疫苗,核酸疫苗具有易操作、低成本等优点,但是其内在危险性还有待进一步研究和排除,在其制备过程中可能掺入的细菌内毒素就是影响因素之一.细菌内毒素是革兰氏阴性菌细胞壁外膜结构中的脂多糖,体内释放后会引起发热、休克等多种病理变化.国外已有体外实验证明内毒素会对小鼠的生殖系统产生影响,本研究旨在通过在体实验探讨内毒素对免疫小鼠的存活率和产仔率的影响.采用常规方法(碱裂解法)、试剂盒2种方法,提取2种pCR3.1-Sry质粒(2种质粒的区别在于是否去除内毒素,其浓度和纯度均经过琼脂糖凝胶电泳和紫外分光光度法分析和控制).作为核酸疫苗对小鼠进行免疫:全程统计免疫小鼠的存活率和产仔率,同时注射灭菌ddH2O作为对照.分子生物学实验结果显示,所制备疫苗具有适宜的浓度和较高的纯度;免疫实验和数字统计结果显示,未去除内毒素实验组小鼠较去除内毒素组的存活率由93%下降到84%、产仔率由87.5%下降到35.7%,而去除内毒素组却与对照组没有明显差异.总之,核酸疫苗制备过程中掺入的细菌内毒素会对免疫小鼠造成较大不良影响,特别是生殖系统,体现在产仔率大幅降低.     

艾滋病疫苗的研究进展

2005年初,在全球六大州19个国家共有35个艾滋病疫苗候选进入了人体早期临床实验。选用的抗原一般均着眼于诱导细胞免疫和体液免疫双重效果。从疫苗的存在形式上看,可供选择的HIV候选疫苗包括下列几种:传统疫苗(灭活疫苗和减毒活疫苗)、合成肽和蛋白亚单位疫苗、DNA疫苗以及活载体疫苗。目前新型疫苗研究策略包括抗原改造加强免疫原性、免疫佐剂协同以提高疫苗免疫反应强度;新型免疫策略包括初免/加强免疫策略和粘膜免疫策略。由于粘膜感染是艾滋病病毒感染的主要途径之一,因而,探索经粘膜免疫是未来疫苗发展的重要方向。中国艾滋病疫苗研究领域的现状是挑战与机遇并存,需要相关领域专家的共同努力,以促进我国艾滋病疫苗研究的发展。     

Humoral and cell-mediated immune responses to live recombinant BCG-HIV vaccines

Several viral and bacterial live recombinant vaccine vehicles are being developed to produce a new generation of vaccines against a broad spectrum of infectious diseases. The human tuberculosis vaccine Mycobacterium bovis bacillus Calmette-Guerin (BCG) has features that make it a particularly attractive live recombinant vaccine vehicle. BCG and other mycobacteria are highly effective adjuvants, and the immune response to mycobacteria has been studied extensively. With nearly two billion immunizations, BCG has a long record of safe use in man. It is one of the few vaccines that can be given at birth, it engenders long-lived immune responses with only a single dose, and there is a worldwide distribution network with experience in BCG vaccination. Recently developed molecular genetic tools and methods for mycobacteria have provided the means to introduce foreign genes into BCG. Here we report that a variety of human immunodeficiency virus type 1 polypeptides can be expressed in BCG recombinants under the control of the mycobacterial hsp70 promoter and that the foreign polypeptides produced in BCG can induce antibody and T-cell responses. These results demonstrate that BCG can be used as a live recombinant vaccine vehicle to induce immune responses to pathogen proteins produced by the bacillus.     

基因疫苗研究与进展

基因疫苗代表了新的、具有潜力的疫苗发展途径和研究方向，它取得成功主要归功于基因治疗技术的发展．基因疫苗是直接将抗原的编码基因注入动物体内，并在动物细胞中表达抗原蛋白以诱导动物产生免疫保护作用．基因疫苗研究有可能为当今人类无法控制的一些严重传染疾病，提供新的免疫防治途径和手段．本文回顾了基因疫苗发展历史和研究进展，与传统疫苗相比较，对基因疫苗的作用原理、构造以及技术方法等方面的初步研究和进展情况进行了概述。     

反祛魅：海派鬼文学的主要特征

海派鬼文学并没有以科学精神祛除鬼文化的幻魅,反之,它包容了传统精神。鬼文学再现了中国鬼文化的神秘,凸显了中国人的怕鬼心理。对鬼的可爱,它也多有涉及,这主要体现在人鬼恋情上。海派鬼文学反拔了现代文学的新传统,继续书写了鬼的奇幻。     

Tumor carbohydrate antigens and strategies to develop cancer vaccines and drugs

Certain carbohydrate antigens of malignantly transformed cells have been identified as markers for the onset of cancer and have become targets for the development of anticancer vaccine therapies. For tumor antigens, many carbohydrate antigens belong to T-independent (TI) antigens. Carbohydrate conjugated to protein carriers can switch TI antigen to a T-dependent (TD) antigen. Attempts to add an innate immune response element (such as Toll-like receptor ligand) to carbohydrate TI-antigens have also been studied. Glycosylation inhibitors or small interfering RNA have also been used for antitumor and/or antiviral agents. This review aims at describing the vast spectrum of tumor carbohydrate antigens and strategies to develop cancer vaccines and drugs.     

pH-responsive vaccine delivery systems for improving cellular immunity

Immune vaccination is a critically important strategy in disease prevention and treatment. In vaccination, efficient vaccine carriers are necessary to augment the immune response initiated by vaccines generally with weak immunogenecity. Nowadays, commercially available vaccine/carrier formulations induce effective humoral immunity but weak or none cellular immunity. However, in practice, cellular rather than humoral immunity plays the key role in treating some intractable diseases such as AIDS and cancers, in which the elicited cytotoxic T lymphocytes can directly kill HIV-infected or cancer cells. To trigger potent cellular immunity, the carriers should help antigens escape from endosomal/lysosomal degradation and release them directly into the cytosol of antigen presenting cells. To this aim, such kind of vaccine carriers should be rationally designed and prepared in terms of their structure and physiochemical properties. Here, we summarized the recent advances in pH-responsive vaccine carriers exclusively developed for improving cellular immunity.     

Successful vaccination with a polyvalent live vector despite existing immunity to an expressed antigen

A global vaccination strategy must take into account production and delivery costs as well as efficacy and safety. A heat-stable, polyvalent vaccine that requires only one inoculation and induces a high level of humoral and cellular immunity against several diseases is therefore desirable. A new approach is to use live microorganisms such as mycobacteria, enteric bacteria, adenoviruses, herpesviruses and poxviruses as vaccine vectors. A potential limitation of live polyvalent vaccines, however, is existing immunity within the target population not only to the vector, but to any of the expressed antigens. This could restrict replication of the vector, curtail expression of antigens, and reduce the total immune response to the vaccine. Recently acquired immunity to vaccinia virus can severely limit the efficacy of a live recombinant vaccinia-based vaccine, so a strategy involving closely spaced inoculations with the same vector expressing different antigens may present difficulties. We have constructed a recombinant vaccinia virus that expresses surface proteins from two diverse pathogens, influenza A virus haemagglutinin and herpes simplex virus type 1 (HSV-1) glycoprotein D. Mice that had recently recovered from infection with either HSV-1 or influenza A virus could still be effectively immunized with the double recombinant.     

A malaria T-cell epitope recognized in association with most mouse and human MHC class II molecules

An ideal vaccine should elicit a long lasting immune response against the natural parasite, both at the T- and B-cell level. The immune response should occur in all individuals and be directed against determinants that do not vary in the natural parasite population. A major problem in designing synthetic peptide vaccines is that T cells generally recognize peptide antigens only in association with one or a few of the many variants of major histocompatibility complex (MHC) antigens. During the characterization of epitopes of the malaria parasite Plasmodium falciparum that are recognized by human T cells, we analysed a sequence of the circumsporozoite protein, and found that synthetic peptides corresponding to this sequence are recognized by T cells in association with many different MHC class II molecules, both in mouse and in man. This region of the circumsporozoite protein is invariant in different parasite isolates. Peptides derived from this region should be capable of inducing T-cell responses in individuals of most HLA-DR types, and may represent good candidates for inclusion in an effective anti-malaria peptide vaccine.     

Prophylactic cancer vaccine,from concept to reality？

The demonstration that for infectious diseasesvaccine-induced immunity is in principle only effectivebefore rather than after infection occurs, provides valuableinsights in understanding the nature of immune system andthe challenges in cancer treatment. Besides the alreadyknown underlying counter-back mechanisms, the astro-nomical numbers of tumor cells in established tumorscould overwhelm the limited amount of specific T cellsinduced by vaccination, which may account for the modestefficiency of immunotherapy against cancer. We speculatethat the long window period for cancer development willallow immune-intervening strategies （e.g., the proper pro-phylactic vaccination） to promote adaptive mechanismstoward an enhanced immunosurveillance, which couldeffectively eradicate or at least control the few precancer-ous cells undergoing neoplastic transformation during earlypremalignant stages in cancer development, and protect thehost from lethal tumor formation. It should be emphasizedthat the pre-cancer-associated antigens but not the tumor-associated antigens seem to be the suitable antigens fordesigning prophylactic cancer vaccines. In addition, anideal prophylactic cancer vaccine may contain multiplepre-cancer-associated antigens, which will provide broadand effective immune protection in a heterogeneous humanpopulation. Finally, we demonstrated that placenta-derivedgp96, which can be readily obtained in high amount forvaccination, has the ability to initiate antitumor T-cellimmunity via association with multiple embryo-cancerantigens. Further understanding placental gp96 associatedwith carcinoembryonic antigen repertoires that orchestrateimmune defense networks against cancer formation willallow to provide an effective prophylactic approach incancer prevention.     

大豆磷脂脂质体与绵羊红细胞膜相互作用的研究(Ⅱ)

用超声法制备大豆磷脂小单层脂质体，低渗溶血法制备绵羊红细胞膜。在一定条件下，将脂质体与红细胞膜温育一定时间后，离心去掉脂质体，用正己烷－异丙醇，提取膜总脂质，用高效液相色谱法分离测定温育前后膜脂组成。结果表明，温育后红细胞膜的磷脂酰胆碱与鞘磷脂物质的量之比（ｎｐｃ／ｎｓＭ）明显增加，这是影响膜流动性的重要参数。上述结果与作者以前用ＤＰＨ荧光偏振技术测定的同样条件下膜流动性变化完全一致。因此，很好地从分子水平上解释了与大豆磷脂脂质体温育后，红细胞膜流动性的增加。     

基于免疫遗传算法的装配顺序优化

采用装配角度、装配方向、装配稳定性和重定位次数4个指标构建装配顺序优化模型,并用疫苗自动获取的免疫遗传算法(IGABVAU)求解.提出4种免疫疫苗用于提升算法运行效率,其中KP和NL型疫苗由人工指定,BP和NT型疫苗在算法运行中自动获取.船舶工段装配实验结果表明,该算法具有较快的收敛速度.进一步研究发现,免疫疫苗的质量会影响疫苗自动获取的免疫遗传算法收敛速度,人工指定的KP和NL型疫苗可以提高初始种群的质量,自动获取的BP和NT型疫苗可以为装配顺序的进化提供方向.