日本血吸虫天然抗病分子疫苗的结构与功能分析 Ⅰ.未成熟卵28kDa分子的快速分离纯化与鉴定

为了进一步研究日本血吸虫天然抗病分子(SIEA 28 kDa)的结构与功能,采用SDS-PAGE制备电泳和电洗脱法从日本血吸虫未成熟卵可溶性抗原(SIEA)中快速分离纯化了SIEA 28 kDa抗原分子,纯化后抗原分子经梯度凝胶电泳分离,重复测定其分子量。同时,采用Western blot和ELISA检测SIEA 28 kDa分子的免疫活性及其生化纯度。证实获得的提取物SIEA 28 kDa抗原分子的纯度高、活性好;用该抗原分子免疫家兔获得的单特异性抗血清可同时识别日本血吸虫未成熟虫卵、雌虫和雄虫的28 kDa抗原组分。说明该分子与日本血吸虫成虫(AWA)28 kDa组分具有部分交叉抗原性。     

ELISA检测姜片虫病患者抗体的研究

研究了姜片吸虫成虫冷浸抗原检测姜片吸虫病患者血清抗体的敏感性和特异性及其在流行病和临床上的应用价值．超声粉碎制备姜片吸虫成虫冷浸抗原．以评价ELISA法检测姜片虫病血清抗体的敏感性、特异性和交叉反应性．按常规ELISA方法操作,目视与酶标仪判断结果．普查2189人中姜片吸虫卵阳性者168例;168例中165例ELISA阳性,ELISA与改良加藤法的阳性符合率98．21％;健康居民血清147份,阳性6份,阳性率4．08％;与血吸虫病交叉阳性为9．38％,肺吸虫病交叉阳性为5．36％．姜片吸虫成虫冷浸抗原1：3500工作浓度包被酶标板,检测姜片虫病血清抗体具有敏感性高、特异强和交叉反应率低的特点．     

大肠杆菌新菌毛抗原的研究V．用标记抗体技术检验菌毛抗原

应用荧光抗体染色、免疫酶染色及斑点酶联免疫吸附试验（Ｄｏｔ－ＥＬＩＳＡ）对大肠杆菌Ｆ１９８７新菌毛抗原进行了检验，经可行性、特异性及对粪便标本检验等试验，表明这些方法均可用于对Ｆ１９８７新菌毛抗原的检出。三种方法除均表现特异、准确外，又分别具有各自的特点。荧光抗体染色结果易分辨，但需具备荧光显微镜；免疫酶染色只需普通显微镜；Ｄｏｔ－ＥＬＩＳＡ可用眼观判定结果，但对病料标本需做一定的处理后才能进行检验。本次试验均采用了间接染色法，同时试验建立了三种方法对Ｆ１９８７新菌毛抗原的具体操作程序。     

乙肝表面抗原小蛋白S基因导入番茄的遗传转化体系的建立

以丽春番茄无菌苗的子叶为外植体，以农杆菌菌株EHA105(包含质粒pBin438S)为载体，将乙肝表面抗原小蛋白S基因导入番茄组织中，基本培养基采用MS 6—BA1．0mg／L IAA0．2mg／L，在共培养时加入乙酰香酮100μmol／L作为诱导剂，经过组织培养得到转基因植株，对转化植株基因组进行的分子学PCR检测表明，外源基因有可能已整合到番茄基因组中，该转基因遗传体系的建立为利用转基因植物生产乙肝口服疫苗奠定了基础。     

大肠杆菌新菌毛抗原的研究Ⅳ．菌株的致病作用检验

应用消化道接种途径，测定了产生Ｆ１９８７新菌毛抗原大肠杆菌菌株对貂、兔、犬、猪、貉子的致病作用，同时对貉子进行了静脉、肌肉接种的感染试验。结果初步表明：供试菌株仅引起了貉子明显发病，经消化道接种能引起腹泻并随粪便向外界排菌，剖检可见出现眼观、组织学及超微结构的病变，且在肠道内容物中能回收到原感染菌。     

Cloning and expression of HIV-1 p24 gene in insect cells by using BAC-TO-BAC system

Recombinant transposing vector pFHIV24 was constructed by cloning the HIV-1 p24 gene into the Multiple cloning site (MCS) of the transposing vector pFastBac1 in the correct orientation with respect to the polyhedrin promoter. Recombinant bacmid bHIV24 was obtained by transposing a mini-att Tn7 element from the recombinant pFHIV24 to the mini-att Tn7 attachment site on the bacmid by Tn7 transposition functions provided by the helper plasmid. Minipreparation of recombinant bacmid DNA was transfected intoSpodoptera frugiperda (Sf9) cells to get the recombinant virus. Fresh insect Sf9 cells were infected with the recombinant virus containing p24 to express the target protein. The target protein expressed was analyzed on a 15% polyacrylamide gels and then used as antigen to check HIV-1 positive serum by ELISA. Our positive result shows that the expressed p24 protein could be used as standard antigen for HIV-1 diagnosis by ELISA and other reliable diagnostic methods of HIV-1 infection. Supported by the World Bank Boan Program Mallam Nock Joshua: born in 1967, Master of Science To whom correspondence should be addressed (027-7882712-2938)     

基因工程菌株产生的含人类绒毛膜促性腺激素抗原的研究 Ⅲ.β—Galactosidase—hCG 免疫和生物活性的初步探讨

基因工程菌株产生的含人类绒毛膜促性腺激素(hCG)β亚单位 C 末端36肽的β-半乳糖苷酶杂交蛋白(简称:β-Galactosidasc-hCG),主动免疫昆明种小白鼠,能诱发特异抗 hCG 抗体的产生。其抗体滴度的高低与免疫原用量有关。通过对小鼠子宫增重抑制试验,表明其抗血清能中和 hCG 的生物活性。     

Endothelial cells as antigen-presenting cells: role in human transplant rejection

The immunological properties of human endothelial cells suggest they perform a pivotal role in acute and chronic rejection following solid organ transplantation. In this review the basic features of acute and chronic rejection are described as are the cellular and molecular requirements for antigen presentation. Traditionally, antigen-presenting cells are considered to be bone marrow-derived cells. However, these conclusions have been derived from rodent models of allograft rejection where bone marrow-derived passenger leukocytes are the only source of donor major histocompatibility complex (MHC) class II in the grafted organ. In contrast, in humans, virtually all the microvascular and small vessel endothelial cells are ‘constitutively’ positive for MHC class II antigens. The phenotypic properties of human endothelial cells, their response to cytokines and their ability to stimulate resting T cells are described. Unlike bone marrow-derived antigen presenting cells (APCs), which utilise B7/CD28 interactions, human endothelial cells utilise lymphocyte function antigen 3 (LFA3)/CD2 pathways to stimulate T cells. They activate a CD45RO + B7-independent subpopulation of T cells. Their effect on allogeneic T cells is compared with other non-bone marrow-derived cells such as fibroblasts, epithelial cells and smooth muscle cells, which are unable to stimulate resting T cells. Evidence is presented suggesting that release of MHC and non-human leukocyte antigens (HLA) from endothelial cells stimulates an alloantibody and autoimmune response leading to chronic rejection. Received 30 March 1998; received after revision 4 May 1998; accepted 4 May 1998     

结核分枝杆菌特异性抗原的N末端氨基酸序列分析

把ＷｅｓｔｅｒｎＢｌｏｔ分析抗原抗体反应的高度特异性与ＰＶＤＦ膜作为载体进行蛋白质氨基酸序列分析的高度敏感性相结合对结核分枝菌的特异性抗原表位刊物筛选和分析鉴定,获得了一个分子质量为３１ｋｕ,Ｎ末端为ＡｌａＧｌｕＶａｌＡｓｐＬｅｕＶａｌＰｈｅＡｌａＶａｌＳｅｒＴｒｐＰｒｏＶａｌＧｌｙ的结核分枝杆菌抗原。     

Microplate chemiluminescence enzyme immunoassay for the quantitative evaluation of carbohydrate antigen 72-4 in human serum

A highly sensitive and specific microplate chemiluminescence enzyme immunoassay （CLEIA） was developed for the quantitative evaluation of carbohydrate antigen 72-4 （CA72-4） in human serum, using luminol-H2O2 catalyzed by horseradish peroxidase （HRP） as the chemiluminescence system. The simple and quick determination was accomplished through a sandwich reaction mode. Several physicochemical parameters of the immunoreaction, including incubation conditions, antibody coating conditions, dilution ratio of anti-CA72-4-HRP conjugate, and chemiluminescence reaction time, were studied and optimized. The proposed method exhibited a linear range of 0-200 U/mL with correlation coefficient and detection limit of 0.9995 and 0.18 U/mL, respectively. The inter-assay and intra-assay coefficients of variation （CV） were both less than 10%. The average recovery of two clinical sera with low and high concentration CA72-4 was 99.3% and 98.7%, respectively. Normal tumor markers, including AFP, CEA, CA24-2, CA19-9 and CA15-3, did not cross-react with each other. The method＇s stability was evaluated by assessing its analytical performance after storing the immunoreagents at 4℃ and 37℃ for 7 days. Little difference was found, indicating satisfactory stability of the method. The present method has been successfully applied to the detection of CA72-4 human serum, and showed a good correlation with the commercially available ELISA kit （r^2=0.9383）. This method showed great potential in the fabrication of diagnostic kit for CA72-4, and could be well used in diagnosis of cancer in clinical practice.