Failure to induce mucosal disease in cattle persistently infected with bovine viral diarrhea virus by treatment with adrenocorticotropic hormone

Recent research has shown that cattle that develop mucosal disease (MD) often, if not always, have been persistently infected with bovine viral diarrhea virus (BVDV) since birth. The purpose of the present study was to determine whether MD could be induced by immunosuppression of persistently BVDV-infected cattle. For that purpose, adrenocorticotropic hormone (ACTH) was injected intramuscularly, twice daily for 5 consecutive days in 4 persistently BVDV-infected cattle and in 3 control cattle. Before the ACTH treatment, the numbers of leukocytes, neutrophils and mononuclear cells (MNC) per litre of blood in BVDV-infected cattle were in the same range as in the controls. Similarly, the proportions of B cells, T cells, monocytes and Fcγ⁺ cells (cells with receptor for the Fc part of IgG) were the same in the 2 groups of animals. On the other hand, the proliferative response to mitogen stimulation of MNC obtained from the control animals was twice as high as the corresponding value of the persistently BVDV-infected cattle.In all animals, ACTH treatment caused increased Cortisol concentrations, leukocytosis, neutrophilia and decreased mitogen-induced lymphocyte stimulation. However, the MNC count and the proportions of B cells, T cells, Fcγ⁺ cells and monocytes remained unaltered. In spite of the immunosuppression, indicated by the decrease in mitogen-induced lymphocyte stimulation. ACTH treatment did not provoke any clinical signs of MD in the persistently BVDV-infected cattle.     

In vitro blastogenic responses of peripheral lymphocytes from marmosets to phytohemagglutinin and to autologous lymphocytes transformed by Epstein-Barr virus

White-lipped marmosets were evaluated for their cell mediated immune (CMI) response to EBV to determine the feasibility of CMI studies in marmoset models for EBV oncogenesis. The mitogen, cell concentrations, the length of incubation period and serum requirements were defined for in vitro lymphocyte stimulation tests. The level of response of each animal was dependent on the concentration of phytohemagglutinin-P (PHA-P) and was independent of cell densities employed. The rate of tritiated-thymidine incorporation by mononuclear cells due to PHA-P increased exponentially between 2–4 days. This test was reproducible for a given batch of PHA-P when the cells were cultured in the presence of 10% heat inactivated fetal bovine serum. The five white-lipped marmosets were seronegative for EBV antigens and did not show lymphocyte stimulation with EBV particles and EBV soluble antigen, but two of these animals exhibited significant stimulation with autologous lymphocytes transformed in vitro by B95-8 virus. Despite the limited amount of blood (3–4 ml) that could be obtained from each animal in a single bleeding, it was possible to perform multiple lymphocyte stimulation assays with the protocol used.     

松科植物提取物对肉仔鸡免疫功能影响的研究

从豫西山区野生针叶松树皮中提取生物活性物质,研究其对肉仔鸡免疫功能的影响。选取180只1日龄Avian肉仔鸡随机分为4组,每组3个重复,每个重复15只。A组为对照组,饲喂基础日粮,B、C、D组分别添加500、1000、4000mg/kg松科植物提取物。在21、49日龄时分别测定免疫器官重量,49日龄时测定淋巴细胞转化率及平均日增重。结果表明,日粮中添加松科植物提取物,可以显著提高免疫器官重量和淋巴细胞转化率(P<0.05)。     

啤酒酵母葡聚糖对断奶仔猪生产性能及淋巴细胞转化率的影响

为了探讨啤酒酵母葡聚糖在断奶仔猪日粮中的适宜添加剂量及其对断奶仔猪细胞免疫功能的影响,本研究进行了2个试验。试验1选用100头(28±2)d断奶的二元杂交断奶仔猪,按单因子试验设计随机分为5个处理,分别饲喂含葡聚糖0、25、50、100mg/kg和200mg/kg的日粮。结果表明:随葡聚糖添加剂量的增加,平均日增重在14￣28 d及0￣28 d呈二次曲线变化(P<0.05)。试验2选用80头(28±2)d断奶的二元杂交断奶仔猪,随机分为2个处理,分别饲喂含葡聚糖0mg/kg和50mg/kg的日粮。在试验的第14天和第28天,每重复取1头仔猪前腔静脉采血,测定外周血淋巴细胞转化率。结果显示,在断奶仔猪日粮中添加50mg/kg葡聚糖提高了仔猪在14￣28d及0￣28d的日增重(P<0.05)。而且,也提高了仔猪在0￣14 d、0￣28 d及28￣35 d的平均日采食量(P<0.05)。但是对淋巴细胞转化率没有影响。结果表明:在断奶仔猪日粮中添加50mg/kg啤酒酵母葡聚糖,可以提高断奶仔猪的生产性能,而且没有性别差异。     

黄芪多糖对雏鸡外周血T淋巴细胞转化功能的影响

将１０８只１日龄伊莎系蛋用公雏均分为３组：一组为对照,其余２组在３日龄时,于背侧颈部皮下分别注射０．２、０．４ｍＬ黄芪多糖注射液（０．０１ｇ／ｍＬ）１次,再分别于７、２１、３５、４９日龄时采用ＭＴＴ比色法及微量全血培养３Ｈ－ＴｄＲ掺入法检测外周血Ｔ淋巴细胞转化率的动态变化。结果表明,黄芪多糖对２１、３５日龄雏鸡Ｔ淋巴细胞转化功能有增强作用,且与剂量有相关性,而对７、４９日龄雏鸡的作用不明显。ＭＴＴ比色法与３Ｈ－ＴｄＲ掺入法的检测结果无显著差异（Ｐ＜０．０５）。     

硫酸化人参总皂苷的制备及其对鸡外周血淋巴细胞增殖的影响

用氯磺酸-吡啶试剂制备了硫酸化人参总皂苷衍生物，采用红外光谱仪检测到制备的硫酸化人参总皂苷衍生物中在1230cm^-1和810cm^-1有硫酸酯键的特征吸收峰。为了初步探讨硫酸化人参总皂苷的免疫学活性．以MTT法测定了人参总皂苷及其衍生物对鸡外周血淋巴细胞增殖的影响。结果表明，150μg／m1人参总皂苷和衍生物均能显著抑制鸡外周血淋巴细胞的增殖（P〈0．05），10μg／ml的衍生物能极显著促进淋巴细胞增殖，与同浓度人参总皂苷相比活性极显著增强（P〈0．01）。实验结果提示，对人参总皂苷进行硫酸化修饰是可行的，经硫酸化后人参总皂苷衍生物的免疫活性明显增强。     

人工感染猪流行性腹泻病毒仔猪外周血SmIg+B细胞及T细胞的动态观察

本实验对猪流行性腹泻病毒(PEDV)—J毒株细胞培养毒接种的6头仔猪和3头健康仔猪外周血SmIg~+B细胞及T淋巴细胞的数量作了检测。T细胞用总E玫瑰花(Et)及活化E玫瑰花(Ea)试验作检测,SmIg~+B细胞用荧光素标记的葡萄球菌A蛋白(FITC—SPA)菌体法鉴定。结果:接种后,SmIg~+B细胞百分率显著上升,于第23天时达峰值(29.19±3.45),明显高于对照猪(P<0.05),Et玫瑰花百分率虽略有上升,但与接种前相比无统计学差异(P>0.05),而Ea玫瑰花百分率则显著低于接种前水平(P<0.01)。以上结果表明,PEDV—J毒株细胞培养毒可刺激仔猪B细胞明显增多。由此认为,B淋巴细胞积极参与PED的免疫反应过程,体液免疫是PED猪的主要免疫反应。     

TCDCA对有丝分裂原刺激的小鼠脾淋巴细胞免疫功能的影响

为了探讨牛磺鹅去氧胆酸(TCDcA)对有丝分裂原刺激的小鼠脾淋巴细胞免疫功能的影响,设计了TCDCA的0.01,0.05,0.1,1,10μg/mL5个剂量浓度梯度,开展了TCDCA对有丝分裂原刺激的小鼠脾淋巴细胞增殖反应、分泌IL-2和IgG的影响研究。结果表明:TCDCA对ConA刺激的脾淋巴细胞增殖反应表现出调节作用,对LPS刺激的脾淋巴细胞无明显作用;TCDCA对ConA和LPS刺激的脾淋巴细胞的分泌IL-2表现出向上抛物线形式的明显的调节作用;TCDCA对LPS刺激的脾淋巴细胞分泌IgG具有向下抛物线形式的明显的调节作用。说明TCDCA对有丝分裂原刺激的小鼠脾淋巴细胞免疫功能具有明显的调节作用。     

连花柴芩可溶性粉对鸡风热犯肺证的临床试验研究

【目的】 研究连花柴芩可溶性粉对风热犯肺所导致的鸡发热、咳嗽、食欲减退、口渴喜饮等证候的临床疗效。【方法】 采用鸡传染性支气管炎病毒人工攻毒建立风热犯肺证肉鸡病例模型,将出现典型风热犯肺证临床症状的150只鸡纳入到试验中,并随机分为5组,30只/组,分别为连花柴芩可溶性粉高、中、低剂量组（连花柴芩可溶性粉浓度分别为4、2、1 g/L）,麻杏石甘颗粒药物对照组（1 g/L）,模型对照组,另设30只为健康对照组。除健康对照组和模型对照组外,各试验组连续饮水给药5 d,记录给药前后鸡的精神状态、体温、呼吸道症状、采食及饮水的变化,并检测各组鸡的免疫器官指数、外周血免疫细胞亚群指标,综合考察连花柴芩可溶性粉的临床疗效。【结果】 连花柴芩可溶性粉高、中剂量组对风热犯肺所致的鸡发热、咳嗽的总有效率分别为90.00%和86.66%,显著高于模型对照组（P<0.05）,且对病鸡采食量下降、饮水量增多具有较好的缓解效果（P<0.05）;连花柴芩可溶性粉能提高病鸡免疫器官指数,促进病鸡外周血中CD3⁺、CD4⁺、CD8⁺ T淋巴细胞增殖,且连花柴芩可溶性粉高、中剂量显著高于模型对照组（P<0.05）。【结论】 连花柴芩可溶性粉对风热犯肺所致的鸡发热、咳嗽、食欲减退、口渴喜饮具有较好的缓解和治疗效果,结合药物量效关系和经济成本等因素综合考虑,推荐连花柴芩可溶性粉临床应用剂量为2 g/L,饮水给药,连用5 d。     

稳定表达山羊SLAM受体的BHK-21细胞系的建立及应用

信号淋巴激活分子(SLAM)又称为CD150,是小反刍兽疫病毒(PPRV)和犬瘟热病毒(CDV)等麻疹病毒属病毒感染淋巴细胞的主要受体,在病毒侵入细胞中发挥重要作用。为建立稳定表达山羊SLAM(g SLAM)的真核细胞系,本研究将人工合成的g SLAM基因克隆至真核表达质粒p IRESpuro3中,并在该基因的3'端引入Flag标签序列作为分子标记,构建了重组质粒p IRES3-g SLAM。将该重组质粒转染BHK-21细胞,经嘌呤霉素加压筛选及采用表达绿色荧光蛋白(GFP)的重组PPRV病毒(r PPRV/GFP)感染鉴定后,筛选到稳定表达g SLAM基因的细胞系。r PPRV/GFP感染和western blot鉴定表明,无论是否有嘌呤霉素压力的存在,该细胞系在传代至第20代,仍能稳定表达g SLAM蛋白。由于g SLAM氨基酸序列与犬的同源性较高,以表达GFP重组CDV强毒株(r CDV/GFP)感染该细胞系,病毒可以感染且能形成明显的细胞病变,表明该细胞系可用于CDV强毒分离和致弱机制等相关研究。