新型免疫佐剂的研究现状及应用

近年来,随着免疫学研究的不断深入及基因工程技术的迅速发展,活载体疫苗、DNA疫苗等新型疫苗的研究也取得了可喜的进步,但这些疫苗免疫原性弱,诱导机体产生的免疫应答不够强,需要配合佐剂以提高其免疫原性。然而常规佐剂存在着一定的局限性,于是新型佐剂的研究受到越来越多的关注。对一些新型免疫佐剂的研究现状和应用前景进行简要的综述和分析。     

疫苗及其在水生动物疾病预防中的应用(四)

正上接2018年9期四、鱼类免疫佐剂的研究及其前景免疫佐剂简称佐剂,是先于抗原或与抗原同时使用时,能非特异性地改变或增强机体产生免疫应答,增强抗原的免疫原性,而本身并不引起机体产生免疫应答,可以积极发挥辅助作用的一类物质。近几年来,随着免疫学研究的不断深入,渔用减毒活疫苗、亚单位疫苗和DNA疫苗等新型疫苗的研究逐步深入,也促进了疫苗佐剂研究的迅速发展。疫苗佐剂作为生物反应调节中的免疫调节剂,可刺激机体产生一定的非特异性免疫反应,且无毒副作用、     

渔用疫苗免疫佐剂应用研究

<正>免疫佐剂简称佐剂,是先于抗原或与抗原同时使用时,能非特异性地改变或增强机体特异性免疫应答、增强抗原的免疫原性,而本身并不引起机体产生相应的免疫应答、可以积极发挥辅助作用的一类物质。近年来,随着免疫学研究的不断深入,渔用减毒活疫苗、亚单位疫苗和DNA疫苗等新型疫苗的研究已逐步开始,也促进了疫苗佐剂研究的迅速发展。与传统疫苗相比,开发新型     

佐剂对DNA疫苗的影响

DNA疫苗是近年来发展较为迅速的一类生物制剂,它能诱导动物机体产生持久的体液免疫和细胞免疫,其在抗病毒、细菌和寄生虫的感染方面起到了重要作用。但与传统的灭活疫苗相比,其免疫效果并不理想。为了攻克DNA疫苗免疫效力低下的难题,一些学者已经研制了多种佐剂制品,以提高DNA疫苗对动物疾病免疫保护的能力。该文针对使用佐剂提高DNA疫苗免疫效力的最新概况做一综述。     

水产疫苗的细胞因子免疫佐剂研究及应用

佐剂（Adjuvant）可通过增强巨噬细胞活性而促进机体T细胞或B细胞的反应，参与半抗原或抗原免疫应答。随着人们对渔用疫苗免疫效果的研究，细胞因子（Cytokine, CK）作为新型佐剂能显著增强疫苗的特异性免疫反应。本文综述了白细胞介素（Interleukin, IL）、干扰素（Interferon, IFN）、肿瘤坏死因子（Tumor necrosis factor-α, TNF-α）、集落刺激因子（Colony-stimulating factor, CSF）、趋化因子（Chemokines）和生长因子（Growth factor）类型、特征及作用及作为渔用疫苗佐剂的研究进展，为系统了解和深入研究渔用细胞因子佐剂的分子机制和实际应用提供参考。     

细胞因子作为分子佐剂在基因免疫中的作用

细胞因子是体内免疫细胞或非免疫细胞产生的一组具有广泛生物学活性的异质性肽类调节因子,在体内能激活和调节免疫活性细胞,对免疫应答的产生和调节具有重要作用,目前应用细胞因子作为免疫佐剂在基因治疗中的研究已有许多报道,本文将IL-2、IL-12和GM-CSF作为分子佐剂在基因免疫中的作用作一综述。     

鱼用免疫佐剂的研究概况

鱼用免疫佐剂配合鱼用疫苗使用是控制鱼类传染性疾病的有效途径,佐剂在鱼病防治中的应用已有了大量研究。从佐剂的类型及在鱼病免疫防治中的研究、作用机理、目前存在的问题和发展趋势几个方面对近年来鱼用免疫佐剂的研究概况作一综述。     

鱼用免疫佐剂的研究概况

鱼用免疫佐剂配合鱼用疫苗使用是控制鱼类传染性疾病的有效途径,佐剂在鱼病防治中的应用已有了大量研究.从佐剂的类型及在鱼病免疫防治中的研究、作用机理、目前存在的问题和发展趋势几个方面对近年来鱼用免疫佐剂的研究概况作一综述.     

草鱼四联佐剂疫苗的制备及特性分析

采用PHA作为非特异性免疫刺激剂与出血病毒、荧光假单胞菌、肠型点状产气单胞菌、鱼害粘球菌4种纯抗原，以最佳方式配合制成四联佐剂疫苗，同时对四联佐剂疫苗的特性进行了分析。结果表明：四联佐剂疫苗的免疫原性极强，防病效果好。     

渔用口服疫苗佐剂研究现状及趋势

正随着水产业的蓬勃发展,人们对渔用口服疫苗的研究不断深入,渔用口服疫苗佐剂也成为研究热点。目前,人们已研制出多种渔用口服免疫佐剂,常见的有聚合微球体、免疫复合刺激物、霍乱毒素、大肠杆菌不耐热肠毒素和细胞因子等。陈昌福等以莨菪碱作为佐剂口服免疫接种草     

鱼用疫苗研究进展

鱼用疫苗的种类繁多，除传统的死疫苗、活疫苗、多价苗之外，新型疫苗包括合成肽疫苗、DNA疫苗、活载体疫苗、基因缺失疫苗等。一些与鱼用疫苗相关的领域如免疫佐剂、接种方法的研究也逐渐深入。本文对以上研究进行了概述，为我国鱼用疫苗的进一步研究和发展提供一些依据。     

二种佐剂对哈氏弧菌OMPC-溶藻弧菌LPS偶联疫苗的增效作用

制备哈氏弧菌SpGY020601外膜蛋白复合物(OMPC)和溶藻弧菌EpGS021001脂多糖(LPS)偶联疫苗,设置偶联疫苗组、偶联疫苗 ISA763佐剂组、偶联疫苗 β-葡聚糖佐剂(DEAE)组以及生理盐水对照组,并以相同程序免疫卵型鲳鲹。溶菌酶活性检测、微量凝集反应试验结果显示:各疫苗组血清中的溶菌酶含量无显著差异(P>0.05),但均远高于生理盐水对照组(P<0.05);ISA763佐剂组抗哈氏弧菌SpGY020601抗体滴度较其他组出现的早,抗溶藻弧菌EpGS021001抗体滴度较其他组持续时间长;β-葡聚糖佐剂组次之,其次是无佐剂组,对照组始终未检测到抗体。各组分为腹腔注射攻毒与浸泡攻毒,对EpGS021001的攻击,ISA763佐剂组相对保护率分别为90%和70%,高于β-葡聚糖佐剂组的85%和55%,以及无佐剂组的80%和55%;对SpGY020601的攻击,ISA763佐剂组相对保护率分别为95%和80%,高于或相当于β-葡聚糖佐剂组的95%和75%,以及无佐剂组的90%和75%。     

不同佐剂对大鲵嗜水气单胞菌灭活疫苗的免疫效果

为了探究添加蜂胶佐剂、弗氏佐剂和铝胶佐剂后对大鲵（Andrias davidianus）嗜水气单胞菌（Aeromonas hydrophila）灭活疫苗的免疫效果,取已制备好的大鲵嗜水气单胞菌灭活疫苗,分别加入3种佐剂,充分混匀后制备成含菌量为1×108 CFU/mL的佐剂疫苗,通过腹腔注射免疫健康大鲵(平均体长40 cm、平均体质量85 g）,每尾1 mL,并设不加佐剂组和对照组;分别于免疫后第1、4、7、14、21、28、35天尾静脉采血,利用凝集反应检测血清抗体效价,于免疫后第35天进行攻毒感染试验。结果表明,与对照组相比,蜂胶佐剂组抗体效价于免疫后第1～14天呈上升趋势,第21天达最高,为1: 469.33,弗氏佐剂组于免疫后第1～28天呈上升趋势,第35天达到最大值,为1: 448.00,随后呈下降趋势;铝胶佐剂组抗体效价于免疫后第1～21天呈上升趋势,第28天达到最大值,为1: 362.67,不加佐剂组于免疫后第21天达最高,为1: 384.00,随后呈下降趋势,而对照组基本稳定在1:4.00。攻毒感染试验表明,蜂胶佐剂组、弗氏佐剂组、铝胶佐剂组和不加佐剂组的死亡率为10%、20%、25%和35%,相对免疫保护率分别为88.9%、77.8%、72.2%和61.1%,对照组大鲵死亡率为90%;3种佐剂疫苗均有较好的免疫保护效果,其中以蜂胶佐剂疫苗效果最佳。     

A review on the recent advances and application of vaccines against fish pathogens in aquaculture

Globally, aquaculture has faced serious economic problems due to bacterial, viral, and various other infectious diseases of different origins. Even though such diseases are being detected and simultaneously treated with several therapeutic and prophylactic methods, the broad-spectrum activity of vaccines plays a vital role as a preventive measure in aquaculture. However, treatments like use of antibiotics and probiotics seem to be less effective when new mutant strains develop and disease causing pathogens become resistant to commonly used antibiotics. Therefore, vaccines developed by using recent advanced molecular techniques can be considered as an effective way of treating disease causing pathogens in aquatic organisms. The present review emphasizes on the current advances in technology and future outlook with reference to different types of vaccines used in the aquaculture industries. Beginning with traditional killed/inactivated and live attenuated vaccines, this work culminates in the review of modern new generation ones including recombinant, synthetic peptides, mucosal and DNA, subunit, nanoparticle-based and plant-based edible vaccines, reverse vaccinology, and monovalent and polyvalent vaccines.

     

草鱼出血病基因疫苗的免疫效果

为探讨含有草鱼呼肠孤病毒外衣壳蛋白VP6基因的重组基因疫苗对草鱼出血病的免疫效果，将重组基因疫苗分别配制为10、30和60μg三个梯度，同时设30μg pFastBacTMDual空载体组，均以等体积的氢氧化铝佐剂为免疫增强剂，对草鱼进行注射，以未经处理的草鱼作为对照组。定期采血测定抗体效价，最后对各组草鱼进行攻毒试验并计算死亡率和免疫保护力。结果显示：注射疫苗的三组草鱼均产生抗体，10μg组效价为1：6～1：11；30μg组为1：9-1：13；60μg组为1：11-1：17，且各组效价均在第28天测定最高，免疫保护率依次为100％、100％和95％，而空载体组和对照组为70％和0％。研究表明，疫苗能够诱导草鱼产生免疫反应，为草鱼出血病核酸疫苗的生产应用和产业化提供了重要的理论依据。     

Use of Modified Live Vaccines in Aquaculture

Vaccination is an important disease management strategy used to maintain human and animal health worldwide. Vaccines developed for aquaculture have reduced antibiotic use in fish production. Original fish vaccines were bacterins (formalin‐killed bacteria) delivered through immersion or injection that induced humoral (antibody) immunity. Next generation vaccines relied on multiple killed antigens delivered with an adjuvant to enhance vaccine effectiveness. Work in the 1990s showed the use of various strategies to develop modified live vaccines for use in fish. A modified live vaccine is a live pathogen that has been rendered non‐pathogenic or avirulent by physical, chemical, or genetic engineering methods. The modified live vaccine typically retains its ability to infect the host which allows for effective presentation of protective antigens to generate cellular immunity (CD4 or CD8 T‐cell responses). Modified live vaccines are advantageous in that they can be easily delivered (i.e., by immersion to young fish) and stimulate both humoral and cellular immunity of long duration. Disadvantages include issues with modified live vaccine safety to the host and environment. A successful modified live vaccine for use in warm water aquaculture is used to highlight the live vaccine strategy.     

Comparison of different challenge methods to evaluate the efficacy of furunculosis vaccines in Atlantic salmon, Salmo salar L.

Four challenge methods, intraperitoneal (i.p.) and intramuscular (i.m.) inoculation, bath and cohabitation exposure, were evaluated as methods for testing the efficacy of furunculosis vaccines in Atlantic salmon, Salmo salar L. Groups of fish vaccinated with one of two different vaccines containing aluminium phosphate or animal/vegetable oil as an adjuvant were challenged with Aeromonas salmonicida 6 and 12 weeks after vaccination. Relative per cent survival (RPS) was calculated daily during a 3-week observation period post-challenge. A large variation in protection measured by RPS, both between methods and between different time points for each method, was found. Towards the end of the observation period, RPS tended to be similar in the i.p. and cohabitation challenge groups. A high degree of protection was demonstrated for the oil adjuvanted vaccine. The i.m. challenge produced very low RPS numbers for both vaccines, but this was most marked for the aluminium-phosphate-adjuvanted vaccine. The bath exposure resulted in RPS values intermediate to the cohabitation and injection methods. The study also demonstrated that, after i.p. and i.m. challenges, the initial peak mortality caused by the inoculation was followed by a secondary increase in mortality, probably because of shedding of bacteria into the water during the first mortality phase and hence contributing to a superinfection state.     

Whole cell inactivated autogenous vaccine effectively protects red Nile tilapia (Oreochromis niloticus) against francisellosis via intraperitoneal injection

Francisella noatunensis subsp. orientalis is a pathogen of tilapia and other warm‐water fish for which no vaccines are commercially available. In this study, a whole cell formalin‐inactivated vaccine was developed for the first time using the highly virulent isolate STIR‐GUS‐F2f7 and the oil‐based adjuvant Montanide? ISA 763A VG. The efficacy of the vaccine was assessed in red Nile tilapia via intraperitoneal (i.p.) injection using homologous experimental infection and correlates of protection such as seral antibody production and bacterial loads in the spleen. For immunization, fish were i.p. injected with 0.1 ml of the vaccine, the adjuvant alone or PBS. At 840 degree days post‐vaccination, all fish were i.p. injected with 4.0 × 10³ CFU/fish of pathogenic bacteria. The RPS at the end of the trial was 100% in the vaccinated group with significantly higher survival than in the adjuvant and control groups. The RPS in the adjuvant group was 42%, and no significant difference was seen in survival between this and the PBS group. Moreover, significantly higher antibody titres in the serum and significantly lower bacterial loads in the spleen were detected in the vaccinated fish by ELISA and qPCR, respectively. These findings highlight the potential of autogenous vaccines for controlling francisellosis in tilapia.