常用新城疫疫苗免疫佐剂的筛选

文章对目前家禽生产中较常见的新城疫疫苗传统免疫佐剂、天然来源佐剂、化学药物佐剂及新型免疫佐剂的种类与作用机理进行综述,并指出疫苗免疫佐剂选择的关键点。     

免疫佐剂作用机理的研究进展

随着DNA重组疫苗、合成肽疫苗等新型疫苗不断涌现,免疫佐剂研究越来越受到人们的关注.尽管佐剂活性物质的研究报道很多,但由于佐剂加强免疫反应是一个非常复杂的过程,关于各种佐剂的作用机制至今尚未完全了解.本文对近年来免疫佐剂作用机理研究进展作-综述,为促进免疫佐剂作用机理的深入研究及免疫佐剂的合理应用提供参考,并初步探讨其作用机理研究的重要性及意义.     

佐剂及其在基因免疫研究中的应用

论述了免疫佐剂的作用机理及细胞因子、CpGDNA、化学佐剂、共刺激信号分子、化学因子、黏附分子等佐剂在基因疫苗中的应用概况，并对免疫佐剂在基因疫苗中的应用前景进行了展望。     

乳剂类免疫佐剂研究进展

乳剂类免疫佐剂又称为油类佐剂，是将抗原包裹在各种油相中，以实现为抗原提供贮存库，并通过将抗原靶向递呈给免疫细胞以增强机体对抗原的特异性免疫应答反应的一类免疫佐剂。自1916年首次报道乳剂类佐剂的应用至今，一百多年来乳剂类佐剂已成为疫苗研制与开发中不可缺少的重要因素，为应对人类和动物各种传染病提供更安全有效的疫苗提供了物质保障。随着现代疫苗学不断的发展，新型疫苗的研制和开发对佐剂提出了更高的要求，了解佐剂的组成成分、有效性和安全性的免疫作用机理是佐剂应用于疫苗研制和开发的前提。因此本文选择了白油佐剂、完全弗氏佐剂(Complete Freund’s Adjuvant,CFA)、不完全弗氏佐剂(Incomplete Freund’s adjuvant,IFA)、MF59^?、AS(Adjuvant systems,AS)系列和Montanides系列等目前常用和商品化的乳剂类免疫佐剂，对其佐剂组成成分、免疫作用机理和临床应用情况等进行综述，以期为乳剂类免疫佐剂的临床使用提供一定的理论参考。     

我国免疫佐剂的研究现状

近年来免疫佐剂研究发展迅猛,新型免疫佐剂层出不穷。介绍了蜂胶佐剂、乳酸杆菌DNA免疫佐剂、弗氏佐剂、脂质体、多糖、中药、硒和维生素E等几种新型免疫佐剂,并简要分析综述了研究新型免疫佐剂的意义和前景。     

复合佐剂的研究进展

不断开发新型免疫佐剂,尤其是复合佐剂,是目前疫苗佐剂的主要发展趋势。根据疫苗生产应用的需要,将单一佐剂复合搭配使用,既可发挥各个佐剂的优点,又有互相促进的作用,达到明显增强免疫效果的目的。为适应生产和市场的需求,复合佐剂对动物机体应无副作用,安全有效。本文对复合免疫佐剂的研究现状和应用前景进行了综述,以期为开发研制高效、低毒的复合免疫佐剂提供参考。     

免疫佐剂的研究进展

佐剂通过调节、增强机体的免疫应答而提高预防和治疗性疫苗的免疫效力。研究开发低毒、高效的免疫佐剂，对疫苗特别是新型疫苗的研发具有重要意义。文中对近年来一些常规佐剂和新型佐剂进行简要综述，为免疫佐剂研制提供有益参考。     

油乳佐剂及其安全性研究进展

油乳佐剂是免疫佐剂的一种,能使多种抗原产生高滴度抗体,是动物疫苗中应用最广泛的佐剂之一.但此种佐剂在发挥佐剂效应的同时,也会来带一些安全性问题,如应激反应、矿物油的残留等.因此,本文拟对油乳佐剂的作用机理、分类、应用以及潜在的安全性问题等作简要综述,并进一步指出有待研究的问题,为后续开展白油佐剂的相关研究提供参考.     

兽用疫苗佐剂的研究进展

兽用疫苗免疫佐剂可以辅助兽用疫苗进入动物机体,增强疫苗的免疫效力,维持疫苗的稳定性,帮助疫苗更加有效地预防、控制动物传染病,减少损失,保障安全的生态环境。用作兽用疫苗免疫佐剂的物质越来越多,有水性疫苗佐剂和油性疫苗佐剂等,各种佐剂的有效成分和作用方式各不相同。本文就兽用疫苗佐剂的特点做了概述,以期为疫苗研发过程中佐剂的筛选提供参考。     

新型兽用疫苗佐剂的研发

现代疫苗大部分是由化学合成多肽或基因工程重组抗原组成的,其免疫原性相对较弱,需要借助免疫佐剂来提高疫苗的免疫效果,具体来讲,免疫佐剂能够提高机体对抗原的适应性免疫应答。现从纳米粒子佐剂、天然佐剂、脂质体、细胞因子分析新型兽用疫苗佐剂的研发和应用。     

Regulating effects of porcine interleukin-6 gene and CpG motifs on immune responses to porcine trivalent vaccines in mice

In order to develop novel immunoadjuvants to boost immune response of conventional vaccines, experiments were conducted to investigate the regulating effects of porcine interleukin-6 gene and CpG motifs as the molecular adjuvants on immune responses of mice that were co-inoculated with trivalent vaccines against Swine fever, the Pasteurellosis and Erysipelas suis. Synthetic oligodeoxynuleotides containing CpG motifs were ligated into pUC18, forming recombinant pUC18-CpG plasmid. Eukaryotic plasmid expressing porcine interleukin-6 (VPIL-6) were also constructed as molecular adjuvants in an attempt to enhance levels of immune responses of mice co-administered with the trivalent vaccines in this paper. The cellular and humoral immune responses of mice were systematically analysed, and the experimental results were observed that the number of white blood cells, monocytes, granuloytes and lymphocytes significantly increased, respectively, in the mice immunized with VPIL-6, compared with those of the control; the IgG content and titre of specific antibodies to the trivalent vaccine mounted remarkably in the sera from the VPIL-6 vaccinated mice; the proliferation of lymphocytes and induced IL-2 activities were significantly increased in the vaccinated groups. The above-mentioned immune responses of mice co-inoculated with pUC18-CpG plasmid were significantly stronger than those of co-inoculated with pUC18 plasmid, suggesting that the immunostimulatory effect of oligodeoxynuleotides CpG is closely connected with the number of CpG motifs. These results suggest that the porcine IL-6 gene and CpG motifs could be employed as effective immunoadjuvants to elevate immunity to conventional vaccines.     

Effect of transgenic expression of porcine interleukin-6 gene and CpG sequences on immune responses of newborn piglets inoculated with Pseudorabies attenuated vaccine

Porcine interleukin-6 gene and CpG sequences were used as immunoadjuvants to enhance the immune responses of newborn piglets to Pseudorabies attenuated vaccine (PAV). The titer of specific antibodies to PAV, the proliferation of lymphocytes and induced IL-2 activities were all examined to identify the immune response of the piglets. The results showed that the immune responses with CpG ODN and porcine interleukin-6 gene were significantly stronger than routine immunities. The data suggests that porcine IL-6 and CpG motifs could be employed as effective immunoadjuvants to raise the humoral and cellular responses of newborn piglets to Pseudorabies attenuated vaccine.     

新型动物疫苗佐剂的研究进展

疫苗佐剂是使疫苗免疫原性充分发挥的工具,目前动物疫苗佐剂主要以铝盐佐剂和油乳佐剂为主。近年来基因重组疫苗和亚单位疫苗发展迅猛,而这些新型疫苗与传统疫苗相比免疫原性较弱,这就对佐剂提出了更高的要求。当前针对佐剂的研究层出不穷,部分佐剂如MF59、AS01、AS03等已经在人用疫苗中成功应用,但应用于动物疫苗还有技术难题需要攻破。蜂胶佐剂目前在动物疫苗中应用较广,且已经占有了一定的市场份额。为充分比较现有新型疫苗佐剂的优缺点,为后续疫苗佐剂的研究提供参考,就目前广泛研究的新型动物疫苗佐剂进行综述。     

免疫佐剂的研究进展

随着DNA重组疫苗、合成肽疫苗等新型疫苗的不断涌现,免疫佐剂研究越来越受到人们的关注.本文综述了近年来免疫佐剂研究概况,着重介绍了目前常用及研究较多的佐剂的特点,并就免疫佐剂现状及研究发展趋势提出自己的见解,为开发研制高效、低毒、结构新颖的免疫佐剂提供参考.     

免疫佐剂研究进展

佐剂的主要作用是提高抗原(免疫原)的免疫原性和免疫反应的可持续性,它能引导机体的免疫系统对抗原产生体液免疫或细胞免疫反应.对佐剂的选择取决于免疫的目的,从用途上分,佐剂可分为试验用佐剂和疫苗用佐剂.前者主要用于特异性抗体的制备,而后者则作为疫苗的必要成分.文章主要介绍目前常用的几种佐剂包括铝盐佐剂、弗氏佐剂、免疫刺激复合物(ISCOM)、脂质体和CpG及其在科研和疫苗中的应用.     

Adjuvants

A number of adjuvants of importance in veterinary vaccines are reviewed. These include mineral salt adjuvants, oil adjuvants, hydrophilic and hydrophobic bloc polymers, hydrocarbons and surface active agents. The current and future applications of these substances and their possible modes of action are discussed. Creation of an adjuvant register by the newly formed European Adjuvant Group should facilitate accumulation of information on adjuvants.     

中药有效成分对动物疫苗的免疫佐剂效果及相关机理研究进展

佐剂作为疫苗的重要组成部分,其自身的安全性及对疫苗的免疫增强效果受到广泛重视。动物疫苗佐剂种类较多,包括铝佐剂、油乳佐剂和脂质体佐剂等。但由于可应用的佐剂有限,疫苗的需求量增大,因此寻求更加安全有效的疫苗佐剂成为新的研究热潮。中药具有易获取、毒性低和免疫活性强等特点,有潜力成为新型疫苗佐剂。笔者对中药有效成分的佐剂活性、联合疫苗后的免疫调节效果及作用机制进行综述,阐明中药有效成分作为动物疫苗佐剂的可能性及研究进展,为新型佐剂的研制提供理论依据。     

Effects of Different Adjuvants on the Immunogenicity of PCV2-Cap Protein

The study was aimed to prepare vaccines with different adjuvants,and research its effects on immunogenicity.The PCV2 Cap gene with its signal peptide removed was connected to pET-28a vector,and then was induced to express,using sodium deoxycholate（DOC）and low concentration of urea to dissolve the inclusion body.Different adjuvants,such as alum-based adjuvants,liposome adjuvants,propolis adjuvants,white oil adjuvants and Freund adjuvant were prepared,together with protein purified to make up subunit vaccines,and commercial inactivated vaccine as positive control,immuning mice,and ELISA method were used to detect changes in concentrations of animal serum antibody and cytokines,evaluated the immune protective effect.Results showed that the expression product in the form of inclusion body,using the DOC could omit dissolving inclusion body protein renaturation steps,and the purification method was simple,the capsid protein obtained had high purity.ELISA assays showed that PCV2-Cap had good immunogenicity.And we found that the water system adjuvants had high immune activity,this could provide important previous experimental data for the commercialization of the subunit vaccine.     

不同佐剂对猪圆环病毒衣壳蛋白免疫原性的影响

试验旨在研究猪圆环病毒不同佐剂疫苗的制备及其对免疫原性的影响。将去除信号肽的PCV2（porcine circovirus type 2,PCV2）Cap蛋白基因连接在pET-28a载体上,进行诱导表达,用脱氧胆酸钠（DOC）和低浓度尿素对表达产物进行溶解,制备氢氧化铝胶体佐剂、脂质体佐剂、弗氏佐剂、白油佐剂、蜂胶佐剂,并与纯化的PCV2-Cap蛋白混合制成亚单位疫苗,以商品化的灭活疫苗作为阳性对照,免疫小鼠,并使用ELISA方法检测动物血清中抗体及细胞因子含量的变化,评估其免疫保护效果。结果显示,表达产物以包涵体的形式存在,使用DOC溶解包涵体可省略蛋白复性步骤,且纯化方法简单,获得纯度较高的衣壳蛋白;ELISA检测结果表明PCV2-Cap蛋白能诱导产生特异性较高的抗体;水系佐剂制备的亚单位疫苗具有较高的免疫活性,为亚单位疫苗的商品化提供重要的数据支持。     

Adjuvant enhancement of humoral immune response to chemically inactivated bovine viral diarrhea virus.

Potentiation of the antibody response to inactivated bovine viral diarrhea virus by immunological adjuvants was studied in guinea pigs and cattle. The inactivated bovine viral diarrhea virus alone was demonstrated to be a weak immunogen. Addition of either 2 mg per mL diethylaminoethyl-dextran or 5% alhydrogel to inactivated bovine viral diarrhea virus did not or only slightly stimulated the antibody response; the combined adjuvants induced a significantly higher titer. A higher concentration (20 mg per mL) of diethylaminoethyl-dextran, on the contrary, suppressed the immune potentiation by the dual adjuvants. Combination of Bordetella bronchiseptica and alhydrogel adjuvants stimulated a high titer of antibody. The titer was further elevated upon revaccination and was significantly higher than that induced by alhydrogel alone. In cattle, alhydrogel enhanced the immune response and the additional inclusion of diethylaminoethyl-dextran did not cause a significant potentiation of the immunity. However, the antibody decay rate was significantly slower when stimulated by the combined adjuvants.