检测胞浆抗原用于急性白血病免疫分型

目的:探讨胞浆抗原在急性白血病免疫分型中的临床意义。方法:用流式细胞仪检测42例初治急性白血病患者胞浆抗原CyCD_3,CyCD_(22),CyCD_(79a)和MPO。结果:T—ALL阳性胞浆抗原主要表达在CyCD_3,阳性率占88.9％。B—ALL阳性胞浆抗原主要表达在CyCD_(79a),AML主要表达CyMPO。结论:CyCD_3和CyCD_(79a),及MPO可分别作为T—ALL,B—ALL和AML系列特异性标志,对正确诊断白血病提供非常有价值的依据。     

Effect of Postnatal Ethanol Exposure on Expression of Differentiation Antigens of Murine Splenic Lymphocytes

Ethanol is a recognized immunosuppressive agent in the chronic alcoholic. However, the effects of ethanol exposure on the developing immune system have not been extensively investigated. This study evaluated the effects of early postnatal ethanol exposure, via breast milk, on splenic lymphocyte differentiation antigen expression in offspring reared by ethanol-fed mice. Maternal mice were fed a liquid diet containing 20% ethanol-derived calories during pregnancy (E-P), pregnancy and lactation (E-PL), or lactation (E-L). Ad libitumfed (C) and pair-fed (PF) control groups, fed a control liquid diet, were included. Expression of differentiation antigens on splenic lymphocytes from 21-day-old offspring reared by females in 1 of the 3 ethanol exposure conditions was evaluated by flow cytometry. Offspring reared by E-P females had similar numbers of splenic lymphocytes as offspring reared by C and pair-fed during pregnancy (PF-P) females. In contrast, offspring reared by E-PL and E-L females had fewer splenic lymphocytes than both PF-PL and PF-L (respectively), and C offspring. The number of Thy 1.2⁺, CD4⁺, CD8⁺, and IgG⁺ (B-cell) splenic lymphocytes was reduced in E-PL and E-L offspring compared with PF and C offspring. E-P offspring had fewer CD4⁺ and IgG⁺ splenic lymphocytes than C, but not PF-P, offspring. The percentage of Thy 1.2⁺ splenic lymphocytes was significantly reduced among E-PL and E-L offspring compared with PF-PL and PF-L (respectively), and C offspring. These results suggest that ethanol exposure of female mice during pregnancy, pregnancy and lactation, or lactation alone, alters the phenotypic development of splenic lymphocytes of offspring reared by these females. The greatest effect on differentiation antigen expression occurred when females consumed ethanol during the period of lactation. We speculate that direct exposure of the nursing offspring to ethanol via the breast milk was responsible for the reductions in specific splenic lymphocyte populations. These data demonstrate that mice reared by females fed ethanol during the early postnatal period have a marked depletion of each of the major subpopulations of splenic lymphocytes, and that Thy 1.2⁺ lymphocytes are differentially sensitive to ethanol.     

蝮蛇毒磷酸二酯酶抗原的制备及性质

为研究蛇类毒腺细胞分化过程中磷酸二酯酶的作用,需制备特异性和敏感性较高的该酶单克隆抗体用于免疫组化研究.本文所介绍为浙江蝮蛇蛇毒中磷酸二酯酶抗原的提取纯化过程,为制备单克隆抗体提供条件.作者采用离子交换技术和分子筛过滤法相结合的方法提取抗原,具有步骤少、分离效率高的特点.所得抗原经聚丙烯酰胺凝胶圆盘电泳呈单一条带,测定分子量为20900.     

活动性结核标志物'H-多肽的实验与临床研究

本题研究是经免疫学途径直接检测人体感染结核菌的情况，为现代结核病的实验诊断、临床监测、流行病调查提供了一个全新的检验指标。作者首先发现了一种仅存在于活动性结核病患者体液中的蛋白成份—活动性结核标志物（ＡｃｔｉｖｅＴｕｂｅｒｃｕｌｏｓｉｓＭａｒｋ—ＡＴＭ）１Ｈ—多肽；并为之创立了独特的检测方法，经四年多临床１９４６０例样本调查中确定了ＡＴＭ的临床价值。将ＡＴＭ检测与ＯＴ皮试、酶联免疫ＥＬＩＳＡ、ＤＮＡ探针、ＰＣＲ基因扩增技术及典型病例组患者行Ｘ线计算机断层摄影（ＣＴ）、磁共振像（ＭＲＩ）等多组对比试验中，实验与临床研究资料分析证明：ＡＴＭ检测的总敏感度为８６．０６％、特异度９６．２４％、准确度９３．４５％、诊断效率为８２．８２％、批内ＣＶ１．２％、批间ＣＶ２．０％、Ｐ＜０．０５。经ＮＭＲ光谱分析结构含有ＣＣＨ２官能团。     

Rational approaches to immune regulation

Our studies are mainly focused on developing strategies of immune regulation. In the case of infectious and neoplastic disease, our approach is to upregulate cell-mediated immunity to viral of tumor antigens using an intracellular bacterium as a vector for targeting these antigens to the major histocompatibility complex (MHC) class I and class II pathways of antigen processing, in addition to exploiting the adjuvant properties of the vector to stimulate innate immunity. In the area of autoimmunity, we are attempting to downregulate the immune response by specific immune intervention directed against autoreactive T cells. In these studies we use murine models for multiple sclerosis. Our approach is to use both rationally designed T cell receptor (TCR) peptide analogs and recombinant viral vectors that express TCR components to regulate the disease.     

Rational approaches to immune regulation

Our laboratory is interested in the properties of proteins that render them immunogenic, and how such immunogenicity may be modulated in vivo. We are attempting to enhance the immune response in the design of more effective vaccines against viral diseases, such as HIV, and against tumor antigens expressed on breast, ovarian, and cervical cancer and B cell lymphomas. Our main approach is to use a facultative intracellular bacterium, Listeria monocytogenes, which has the unusual ability to live and grow in the cytoplasm of the cell and is thus an excellent vector for targeting passenger antigens to the major histocompatibility complex (MHC) class I pathway of antigen processing with the generation of authentic cytotoxic T lymphocytes (CTL) epitopes. In the field of tumor immunotherapy, we are also developing nonliving vaccine vectors for tumor antigens.     

Complement-dependent synergistic effects of rat monoclonal IgG antibodies in vivo

We describe studies aimed at maximizing the effector mechanisms responsible for eliminating target erythrocytes from the circulation in a fully homologous opsonization system in vivo. The effects on the subsequent fate of target erythrocytes were examined in both normal and decomplemented rats preinjected with a variety of rat IgG monoclonal antibodies (mAb) directed against different epitopes on the RTlA^a, the classical class I major histocompatibiliy complex antigen of the DA rat. In general, the clearance of both DA and (DA × PVG)F₁ erythrocytes in normal rats preinjected with various pairs of noncompetitive mAb was very rapid when compared with the overall clearance patterns seen with individual antibodies. With all mAb combinations containing IgG2b or IgG2a, an intact complement system was an essential requirement for augmenting the initial clearance and promoting hepatic sequestration of these target cells. The removal of (DA × PVG)F₁ erythrocytes, expressing half as much antigen, was considerably slower than the DA cells for each antibody pair tested although a notable degree of heterogeneity was observed in the overall behavior of both types of target cells with different mAb combinations. Our results suggest that the limiting effects of low antigen density on the target cells combined with the use of mAb of an isotype like the rat IgG2a can be overcome using pairs of mAb that recognize different epitopes on the same target antigen.     

检测HSV等5种病原体抗体的微阵列技术

目的 建立抗原微阵列技术检测多种病原体抗体。方法 将病原体的基因工程抗原，以电脑操纵的机械手将其点在赖氨酸修饰的玻片上．制备抗原微阵列，与血清反应后随之与Cy3标记的二抗反应，用激光共聚焦扫描仪扫描玻片，检测血清中相应病原体的IgG，并进行了灵敏度和重复性的检测。结果 5种抗原的最低检测限度为HSV1-gG 1．56μg／ml，HSV2-gG 3．125μg／ml，CMV-p150 1．56μg／ml，RV-E1E2 3．125μg/ml，MT-EAM 31．26μg／ml；抗原微阵列与ELISA检测的结果相比有较高的符合率；检测IgG和IgM的灵敏度分别为50pg、10pg；不同批次2张玻片间总体变异系数为IgG 6．52，IgM 18．89。结论 抗原微阵列可用于平行检测血液中病原体特异性抗体。