Characterization of bovine ileal epithelial cell line for lectin binding,susceptibility to enteric pathogens,and TLR mediated immune responses

In this study, primary and immortalized bovine intestinal epithelial cells (BIECs) were characterized for the expression of surface carbohydrate moieties. Primary BIEC-c4 cells showed staining greater than 90 % for 16 lectins but less than 50 % staining for four lectins. Immortalized BIECs showed significantly different lectin binding profile for few lectins compared to BIEC-c4 cells. BIEC-c4 cells were studied for infectivity to E. coli, Salmonella enterica, bovine rotavirus, bovine coronavirus, and bovine viral diarrhea virus. Bovine strain E. coli B41 adhered to BIEC-c4 cells and Salmonella strains S. Dublin and S. Mbandaka showed strong cell invasion. BIEC-c4 cells were susceptible to bovine rotavirus. LPS stimulation upregulated IL-10, IL-8, and IL-6 expression and Poly I:C upregulated TLR 8 and TLR 9 expression. This study provides important knowledge on the glycoconjugate expression profile of primary and immortalized BIECs and infectivity and immune responses of primary BIECs to bacterial and viral pathogens or ligands.     

不同方法构建磺胺二甲氧嘧啶免疫抗原的比较研究

为探讨合成磺胺二甲氧嘧啶(SDM)免疫抗原的有效途径,采用重氮偶合法和戊二醛法分别合成SDM免疫抗原,并对其结构特征、光谱特征和SDM结合比进行比较.结果表明,在合成SDM-BSA中采用重氮偶合法比戊二醛法好.     

斑点免疫吸附试验诊断羊脑多头蚴病的研究

以原头节可溶性粗抗原经Sephadex G-200层析纯化抗原为包被抗原，兔抗羊IgG-HRP结合物为显色剂，建立检测羊脑多头蚴病血清抗体的Dot-ELISA，并以ELISA作平行对照。结果，粗抗原和层析纯化抗原检测86头份羊脑多头蚴病阳性血清，其敏感性分别为94．18％和93．02％，两种抗原的敏感性无显著差异；检测122头份绦虫蚴病阴性血清，18头份棘球蚴病阳性血清，35头份细颈囊尾蚴病阳性血清，其特异性分别为90．29％和95．43％。两种抗原的特异性差异显著。Dot-ELISA和ELISA两种方法的符合率为100％。层析纯化抗原比粗抗原的特异性有了明显提高，而敏感性没有降低。层析纯化抗原和操作术式都具有良好的重复性，Dot-ELISA和ELISA对比试验结果相近，且具简便、快速及不需要复杂设备等优点，是一种检测羊脑多头蚴病血清抗体的理想方法。     

沙门氏菌鞭毛蛋白属共同抗原表位的定位

用淋巴细胞杂交瘤技术研制成４株沙门氏菌属特异单抗ＣＢ８，ｄｅ７，ｃｃ６，ＤＤ４，在免疫转印试验中，证实它们所识别的抗原表位位于鞭毛蛋白上，ＥＬＩＳＡ相加试验及竞争试验显示，ＣＢ８和ｃｃ６与ｄｅ７，ＤＤ４识别的抗原表位不同，表明鞭毛蛋白的属共同抗原具有多种表位的特性。用胰蛋白酶消化鞭毛蛋白，ＳＤＳ－ＰＡＧＥ结果出现两条新带，分子量分别为４２０００（Ｆ４２），２７０００（Ｆ２７），长时间消化，最终只剩下Ｆ２７条带，它可抵制该酶的消化。对该酶切样品的免疫转印结果表明，沙门氏菌鞭毛蛋白属共同抗原表位出现于Ｆ４２和Ｆ２７上，而相应的沙门氏菌分型Ｈ抗原单抗或因子血清识别的表位也在Ｆ４２和Ｆ２７上都得到了证实。这为沙门氏菌鞭毛蛋白抗原的多样性及结构和功能研究提供了证据。     

牛流行热病毒灭活疫苗免疫后牛外周血淋巴细胞亚类分布的研究

对牛流行热病毒灭活疫苗免疫并攻每后，牛外周血淋巴细胞亚类的变化进行了研究。结果表明，灭活苗免疫后，牛的CD4^ 显著升高，可能与参与辅助B淋巴细胞合成抗体有关，攻毒后，γδT细胞均显著上升，免疫组在攻毒后3周仍保持在相当的高水平，IL－2Rα阳性细胞在攻毒后高热期也有显著升高，而CD8^ 细胞没有明显变化。     

禽白血病病毒J亚群(ALV-J)的攻毒试验

将禽白血病病毒 J亚群 (AL V- J)内蒙株 NM876 1和 NM9996人工接种于 1日龄爱维茵肉种鸡 ,通过眼观、病理组织学检查观察了攻毒鸡的病理学特征 ;通过 PCR和 EL ISA技术检测了病毒感染率、抗体变化规律以及病毒和抗体的相关性。结果显示 ,攻毒鸡从第 3周开始即可检出病毒 ,NM876 1株和 NM9996株病毒感染率分别为 71.4 %和6 4 .3% ;从第 5周开始 ,出现较明显的病理学变化 ,病变特征以骨髓、肝脏、心脏、脾、卵巢等组织内髓细胞增生为主 ,而法氏囊、脑、坐骨神经无变化 ;攻毒鸡抗体消长变化有一定的规律 ,一般在 7～ 8周龄和 18～ 2 0周龄时分别出现 1次高峰 ,而在 4～ 6周龄、10周龄和 2 3周龄分别出现 1次低谷 ,提示鸡场进行 EL ISA检测时要避开这一时期 ;攻毒鸡产生耐受性病毒血症 ,即病毒阳性而抗体阴性 (V A- )的比例较高 (7/14 ,9/14 )。通过以上的研究证明 ,AL V- J内蒙株疾病模型复制成功 ,NM876 1、NM9996可作为原型株进行相关研究     

2型猪链球菌的血清学鉴定

本试验首次证实了２型猪链球菌在我国的存在，采用玻片凝集、琼脂扩散及毛细管沉淀等３项试验，对１９９０年以来广东某猪场断奶仔猪暴发流行的败血症、脑膜炎及关节炎病猪中分离的１５株链球菌进行了血清学鉴定。结果表明：１５株链球菌均为２型猪链球菌；用高压法提取抗原进行沉淀反应，其结果比酸热法提取抗原具有简便、敏感、特异性强的特点。     

用酶联免疫吸附试验检测鸡白血病以及控制、根除鸡白血病措施的探讨

用酶联免疫吸附试验(ELISA),对2群鸡的鸡蛋清和1株鸡的马立克氏病疫苗进行检测,发现2群鸡的鸡蛋清中,鸡白血病病毒的阳性率分别是11％和29％,鸡马立克氏病冻干苗隐藏鸡白血病病毒群体特异性(gs)抗原的阳性率为100％。本文指出我国禽苗可能带有鸡的白血病病毒,分析讨论了鸡白血病病毒的垂直传递和水平传播的规律,提出在曾祖代和祖代鸡群中,采用ELISA试验,检测鸡蛋清,能减少以至根除鸡的白血病。     

Analysis of the distribution and clonality of TCR Vβ T cells in cord blood

AIM:To investigate the distribution and clonality of TCR Vβ subfamily T cells in cord blood. METHODS:The CDR3 of TCR Vβ 24 subfamily genes were amplified in mononuclear cells from 13 cases of cord blood. To observe the usage of TCR Vβ repertoire, the PCR products were further labeled with fluorescent and analyzed by genescan technique for the CDR3 size, to evaluate clonality of the detectable TCR Vβ T cells. Peripheral bloods from 10 cases of normal individuals and T cell line Molt-4 and Jurkat served as controls. RESULTS:Only 38.78%±16.26% of 24 Vβ subfamily T cell were selectively expressed in cord blood, predominantly in Vβ 3, 5, 8, 9 and 13, whereas all 24 Vβ subfamilies could be detected in T cells from peripheral blood of normal individuals. Genescan analysis showed that all PCR products of TCR Vβ subfamilies from cord blood or normal individual peripheral blood displayed multi-peaks. CONCLUSION:Some TCR Vβ subfamily T cells were absent in cord blood. All TCR Vβ subfamily T cells in cord blood displayed polyclonality.     

Inhibitory effect of berberine on the activation and proliferation of T lymphocytes

AIM: To investigate the effect of berberine (Ber) on the activation and proliferation of T lymphocytes and its mechanism of action. METHODS: Whole peripheral blood from normal subjects was stimulated with phytohemagglutinin (PHA) or phorbol ester (PDB) plus ionomycin (Ion) and the expression levels of CD69 and CD25 were evaluated with flow cytometry after the staining with appropriate fluorescent monoclonal antibody. The distribution of cell cycles was analyzed by propidium iodide staining and dead cells by 7-aminoactinomycin live staining. RESULTS: 100 μmol/L and 50 μmol/L of Ber had significant inhibition of the expression of CD69 on T cells stimulated with PDB plus Ion or PHA, while effect of 25 μmol/L Ber was not significant. And as time of action extended, the extent of inhibition decreased. For the expression of CD25, Ber at the concentrations as above all exerted significant inhibitory effect in a dose-dependent manner. Moreover, Ber could block lymphocytes cell cycle progression from G₀/G₁ phase to S and G₂/M phase without phase specificity. Besides, live staining analysis revealed that Ber did not have significant cytotoxicity on lymphocytes. CONCLUSIONS: Ber significantly inhibits the expression of early and mid activation antigens of T cells and also blocks the progression of lymphocytes cell cycles. These results suggest that Ber exerts immunosuppression effect through inhibiting the activation and proliferation of T cells.