CpG-ODN佐剂在猪口蹄疫疫苗免疫增强的应用

本研究将CpG佐剂同高端口蹄疫疫苗混合后免疫生长猪,经过液相阻断法对抗免疫后的体效价进行评估。结果表明,CpG佐剂同高端口蹄疫疫苗同时使用可以提高免疫抗体水平和免疫合格率。     

黄芪多糖复配辛伐他汀免疫佐剂对鸡免疫H9N2禽流感疫苗免疫效果的影响

为了实现免疫增强功能优化,将免疫增强剂黄芪多糖和辛伐他汀进行复配,并进一步制备出水包油型和油包水型2种复配油乳佐剂。与灭活H9N2禽流感疫苗联用,颈部皮下接种10日龄海兰褐蛋公鸡,并于二免后定期检测多项免疫指标。结果：通过均质乳化法在合适的条件下能制备出稳定的复配油乳佐剂,且水包油型流动性更佳;接种水包油型灭活H9N2油乳佐剂疫苗后,与对照组相比,血凝抑制抗体效价得到极显著提升,血清IgG抗体含量极显著增加(P<0.001),免疫脏器指数显著升高(P<0.05)。综上,黄芪多糖复配辛伐他汀的免疫佐剂能从多方面提升蛋鸡免疫功能,且水包油型的油乳佐剂效果优于油包水型的油乳佐剂。     

蜂胶作为动物疫苗佐剂的研究及应用

<正>蜂胶(Propolis)是一种具有芳香气味的胶状固体物,由蜜蜂从树干或植物芽孢上采集树脂(树胶)后,与其上腭腺、蜡腺自身分泌的的分泌物混合加工而成。其内含有丰富的生物活性物质,具有多种生物学特性,兼具广谱抗病毒、抗细菌、抗霉菌、消炎、止痒、增强免疫、抗氧化、降血糖、降血脂等多种功能,广泛应用于人类的医疗、保健行业,作为新兴的保健品也倍受推崇,现已成为各国科学研究的热点[1]。蜂胶用作疫苗佐剂优势明显,获得了良好效果,迅速成为了当前佐剂开发热潮中的佼佼者,被广泛应用于动物疫苗领域,并     

卵黄抗体五种提取工艺比较及初步应用

为了得到较优的卵黄抗体提取工艺,试验改良了辛酸法、水稀释法、氯仿法、硫酸铵沉淀法与聚乙二醇沉淀法5种提取方法。IPGT诱导表达重组猪致病性大肠杆菌(Escherichia coli)毒素STxB蛋白(rSTxB),与佐剂71VG乳化制备免疫原免疫蛋鸡,利用改良的聚乙二醇沉淀法提取高免卵黄抗体,采用SDS-PAGE电泳检测抗体纯度,通过Western blot检测卵黄抗体和血清抗体效价。结果表明,辛酸法、水稀释法、氯仿法、硫酸铵沉淀法、聚乙二醇沉淀法提取效率分别为96.00、81.99、155.50、65.34、46.00 mg/mL,其中聚乙二醇沉淀法的提取纯度最高。重组STxB蛋白分子量大小约25 kDa,血清中抗体效价最高达到64 000倍,卵黄抗体最高稀释倍数为16 000倍。综合评估,聚乙二醇的提取法相对较优,且提取的卵黄抗体具有良好的免疫活性。     

新型国产白油佐剂与进口Marcol-52白油佐剂配制疫苗的比较试验

The Newcastle disease inactivated vaccines were prepared with oil adjuvants respectively from domestic and import.The experiments were done in order to compare the new domestic oil and Marcol-52 according to a series of testing indexes. The results showed that the quality of the new domestic oil was better than Marcol-52, except the absorptive quality.     

Preparation of Adjuvant Diluent for Live Vaccine against Classical Swine Fever and Evaluation of its Immune Effects

The aim of this study was to develop a novel oil-in-water (O/W) emulsion as adjuvant diluents (AD) for live vaccine against classical swine fever (CSF) that could effectively enhance the immune effect of vaccine.The AD was prepared by high-pressure homogenization technique.Formulations and preparation parameters were optimized with response surface design.Its stability, particle size, polydispersity (PDI) and Zeta potential were characterized.The humoral immune response and cellular immune response of the AD were evaluated with BALB/c mice by intramuscular injection.The particle size of the AD prepared by optimized formulation and parameters was 100.4 nm, PDI was 0.147, and Zeta potential was —28.7 mV.The experiment results showed that the AD had good stability.The AD was inoculated combined with live vaccine against CSF into BALB/c mice by intramuscular injection.The results showed that the live vaccine against CSF specific immune responses could be evoked in mice by co-inoculation with AD and vaccine.The cellular immune response levels in co-inoculated groups were significantly higher than control group (P<0.05), with obvious phenomena of higher levels of IFN-γ, IL-6 and IL-4 in serum.The result revealed that cellular immune capability significantly improved with the AD.The results strongly revealed that cellular immune capability significantly improved by introducing AD for effective immune-adjuvant for live vaccine against CSF.     

细菌鞭毛蛋白作为佐剂的研究进展

细菌鞭毛蛋白作为Toll样受体5(TLR5)或NOD样受体C4(NLRC4)的配体,其结构决定了其既有抗原性又具有佐剂效应。将细菌鞭毛蛋白与外源抗原混合或融合表达,已获得多种有效的候选疫苗。细菌鞭毛蛋白佐剂效应主要是通过TLR5和NLRC4信号途径协同实现的。TLR5位于细胞表面,可触发炎性因子、趋化因子和Ⅰ型干扰素的分泌,启动天然免疫应答。NLRC4是细胞质中的模式识别受体,可识别细胞质中的多种配体并诱导相应免疫应答。另外,细菌鞭毛蛋白能募集T淋巴细胞和B淋巴细胞至次级淋巴器官,促进DCs和T淋巴细胞的活化。细菌鞭毛蛋白的可塑性将会使得以细菌鞭毛蛋白为基础的疫苗成为当前和未来研究的热点。     

Quil A与PICKCa的免疫调节作用研究

为了探讨Quil A、PICKCa及二者联合使用对实验动物的免疫调节作用,试验分别给小鼠和猪注射Quil A、PICKCa及两者混合物,于注射后不同时间采血,检测细胞因子水平;注射后第7天检测猪T淋巴细胞的体外增殖反应。结果表明:小鼠和猪在注射药物之后均未出现任何不良反应;试验组动物的细胞因子水平均显著高于对照组;猪T淋巴细胞体外增殖能力也显著高于对照组。说明Quil A、PICKCa对小鼠和猪具有良好的免疫调节作用。     

鸡新城疫油佐剂灭活疫苗的研究

采用ＮＤ Ｌａｓｏｔａ株为种毒，福尔马林为灭活剂，７号白油为佐剂，吐温８０、司本８０为乳化剂，硫柳汞为防腐剂，高压匀浆泵为乳化工具，研制出ＮＤ油佐剂灭活疫苗。其剂型为双相型（Ｗ／Ｏ／Ｗ），呈稀薄乳状液，粒度为１ｕｍ左右，粘度≤１ｓ，通针性好。４￣８℃有效保存期１年。效力≥７０ＰＤ５０，最早产生免疫时间为１２天（１００％保护），生产中常用免疫剂量为２月龄内每只０．５ｍｌ，２月龄以上每只１．０ｍｌ皮下     

猪圆环病毒核酸疫苗pcDNA-PCV-ORF2-ISS的构建及分子佐剂联合免疫研究

用PCR方法扩增猪圆环病毒PCVwhn株编码CAP蛋白的基因ORF2,同时人工合成一段CpG序列,并设计上下游引物,进行扩增,定向插入到真核表达载体pcDNA3.1(+)中,构建新型核酸疫苗(pcDNA-PCV-ORF2-ISS),并进行了酶切和测序鉴定.用构建的pcDNA-PCV-ORF2-ISS质粒以及pcDNA-PCV-ORF2与5种细胞因子分子佐剂质粒(IL-6/4、IL-2、IL-4、IL-6、IFN-γ真核表达质粒)联合对仔猪进行免疫试验.免疫后用间接ELISA法测定猪圆环病毒Ⅱ型特异性抗体IgG,用流式细胞仪测定猪血液CD3+、CD4+和CD8+T细胞亚类的数量.结果显示,成功构建含CpG免疫刺激序列的猪圆环病毒Ⅱ型核酸疫苗;pcDNA-PCV-ORF2-ISS在仔猪免疫实验能诱导细胞免疫并产生特异性抗体(P<0.05);用5种细胞因子分子佐荆质粒与pcDNA-PCV-ORF2疫苗同时免疫能显著提高DNA疫苗的免疫效果(P<0.05).分子佐剂免疫增强效果次序为:IL-6、CpG、IL-6/4、IL-2、IL-4和IFN-γ,表明IL-6和CpG最佳.