DNA疫苗的免疫佐剂

本文综述了目前应用于DNA疫苗的几种免疫佐剂,诸如细胞因子、蛋白质分子、CpG寡核苷酸、化学分子、脂质体等,总结了DNA疫苗免疫佐剂的研究现状,探讨了该领域未来可能的发展方向。     

DNA疫苗及其安全性

ＤＮＡ疫苗是在传统疫苗基础上发展起来的一种全新的疫苗，同传统的疫苗相比具有多方面的优越性。随着ＤＮＡ疫苗的深入研究，其应用也越来越广泛。本文回顾了ＤＮＡ疫苗的发展史，论述了ＤＮＡ疫苗的免疫机理和在病原微生物中的防制现状，并探讨了其安全性及需要解决的问题。     

禽类DNA疫苗研究进展

新型疫苗的研究对于禽类传染病的防制意义重大。传统疫苗是基于抗原刺激机体产生特异性抗体的原理,它们大多数激发机体的体液免疫,很难启动细胞免疫。脱氧核糖核酸(DNA)疫苗作为第3代疫苗,具备传统疫苗和其它基因工程苗不可比拟的优点,能够诱导全方位的免疫反应且使用更安全更方便,DNA疫苗是将外源基因与真核质粒重组后直接导入细胞内,使外源基因在宿主细胞内表达合成保护性抗原蛋白。这是模拟病毒自然感染提呈过程,既能产生细胞免疫,又能产生体液免疫。文章对DNA疫苗在禽类应用的可行性和应用研究新进展作了综述。     

减毒沙门菌在禽用疫苗应用中的研究进展

减毒沙门菌感染宿主细胞后,可诱导机体产生强烈的体液免疫、细胞免疫和黏膜免疫反应,不仅可以用作疫苗,而且也是理想的疫苗载体.文章对禽用疫苗中的减毒沙门菌活疫苗、减毒沙门菌携带的重组疫苗和DNA疫苗的研究进行了综述,以期为新型禽用疫苗的研究提供参考.     

猪流感疫苗研究进展

猪流感是由猪流感病毒引起的一种急性高度传染性疾病,可引起病猪高热、食欲不振、呼吸困难、孕猪流产、延迟出栏等,同时该病常继发其他细菌病感染,严重危害养猪业的发展.由于该病具有重要的公共卫生意义,因此研制安全有效的疫苗尤其重要.目前对猪流感疫苗的研究热点主要集中在表位疫苗和利用反向遗传构建弱毒疫苗.论文针对国内外猪流感疫苗的研究情况做了系统描述,包括灭活疫苗、弱毒疫苗、DNA疫苗、载体疫苗等,旨在为新型疫苗的研制提供参考.     

DNA疫苗在动物医学中的应用研究进展

DNA疫苗是继减毒疫苗，灭活疫苗，亚单位疫苗和重组多肽疫苗之后的又一新型疫苗，比其它疫苗高效。安全且易于大量生产。本文对DNA疫苗在牛，猪和鸡一些病毒病上的使用效果及其安全问题等方面的研究近况作一综述。     

DNA疫苗在鱼类中的应用研究进展

DNA疫苗是继减毒疫苗、灭活疫苗、亚单位疫苗和重组多肽疫苗之后的又一新型疫苗,比其它疫苗高效、安全且易于大量生产。本文主要对鱼用DNA疫苗的构建、作用机制、使用方法、使用效果及安全问题等方面研究近况作一综述,以促进我国对鱼用DNA疫苗进一步的研究和应用,从而加速我国水产养殖业的发展,并为深入研究和掌握鱼类的免疫防御机制提供一些依据。     

牛传染性鼻气管炎基因工程疫苗研究进展

牛传染性鼻气管炎是由牛疱疹病毒1型感染而引起的高传染性疾病。该病临床症状以呼吸道炎症为主,另外伴有结膜炎、流产、脑炎等并发症。20世纪50年代早期,美国科罗拉多州某些牧场首次发现该病。目前缺乏有效的治疗性药物,对动物实施疫苗免疫是防控该病较理想的方法。目前,某些国家仍然使用传统疫苗,然而,经过基因改造的糖蛋白E基因缺失疫苗已在欧盟上市,该疫苗分为活苗和灭活苗两种。文章着重对牛传染性鼻气管炎基因缺失疫苗、亚单位疫苗、活载体疫苗及DNA疫苗最新的研究进行评述。     

Toxoplasma gondii surface antigen 1 (SAG1) as a potential candidate to develop vaccine against toxoplasmosis: A systematic review

Toxoplasma gondii is an intracellular parasite that infects a broad range of animal species and humans. As the main surface antigen of the tachyzoite, SAG1 is involved in the process of recognition, adhesion and invasion of host cells. The aim of the current systematic review study is to clarify the latest status of studies in the literature regarding SAG1-associated recombinant proteins or SAG1-associated recombinant DNAs as potential vaccines against toxoplasmosis. Data were systematically collected from six databases including PubMed, Science Direct, Web of Science, Google Scholar, EBSCO and Scopus, up to 1st of January 2019. A total of 87 articles were eligible for inclusion criteria in the current systematic review. The most common antigens used for experimental cocktail vaccines together with SAG1 were ROP2 and SAG2. In addition, the most parasite strains used were RH and ME49. Freund’s adjuvant and cholera toxin have been predominantly utilized. Furthermore, regarding the animal models, route and dose of vaccination, challenge methods, measurement of immune responses and cyst burden have been discussed in the text. Most of these experimental vaccines induce immune responses and have a high degree of protection against parasite infections, increase survival rates and duration and reduce cyst burdens. The data demonstrated that SAG1 antigen has a high potential for use as a vaccine and provided a promising approach for protecting humans and animals against toxoplasmosis.     

犬细小病毒核酸疫苗、重组活载体疫苗与弱毒疫苗免疫犬实验研究

分别用犬细小病毒（CPV）核酸疫苗（pVCPV-VP2）、CPV重组活载体疫苗（CAV2／CPV）与CPV弱毒疫苗对犬进行了免疫试验,以检测不同CPV疫苗的免疫原性。采用CPVELISA、CPVHI与CPV微量中和试验检测免疫犬的体液免疫水平,采用淋巴细胞转化试验检测犬的细胞免疫水平。结果,pVCPV—VP2和CAV2／CPV均能诱导机体产生抗CPVELISA抗体与抗CPV中和抗体,但是pVCPV-VP2不能诱导机体产生可检测的抗CPVHI抗体,而CAV2／CPV能够诱导机体产生抗CPVHI抗体。淋巴细胞转化试验结果,pVCPV-VP2和CAV2／CPV免疫犬的外周血淋巴细胞对ConA与CPV的刺激均出现明显的增殖反应。结果表明,pVCPV—VP2和CAV2／CPV免疫犬均能诱导机体产生抗CPV的特异性体液免疫反应和细胞免疫反应,两者所表达的VP2蛋白均具有较好的免疫原性。CAV2／CPV以及pVCPV—VP2和CAV2／CPV联合免疫犬的抗CPV体液免疫水平和细胞免疫水平均比用pVCPV—VP2单独免疫犬的体液免疫水平和细胞免疫水平高。但CAV2／CPV诱导机体产生的抗CPV特异性免疫反应仍然比CPV弱毒疫苗诱导机体产生的抗CPV特异性免疫反应弱。另外,CAV2／CPV还能诱导机体产生抗CAV-2的特异性免疫反应。     

奶牛乳腺炎金黄色葡萄球菌疫苗研究进展

奶牛乳腺炎是由多种因素引起的严重影响奶牛业发展的重要疾病,金黄色葡萄球菌是引起奶牛乳腺炎的主要病原,至今还没有有效的方法预防金黄色葡萄球菌所致的乳腺炎.虽然已经研制了许多金黄色葡萄球茵疫苗,但是试验表明,这些疫苗的免疫效果并不理想,因此仍将继续搜索有效疫苗.目前,亚单位疫苗和DNA疫苗成为研究热点,研究集中在金黄色葡萄球茵的表面抗原上.利用金黄色葡萄球茵分泌的免疫原性蛋白而制成的疫苗可能具有高免疫效力,这些抗原将成为高效疫苗的候选者.同时,应用低毒的金黄色葡萄球菌突变株,也可能成为预防乳房炎的一种新工具.     

细胞因子类佐剂在核酸疫苗中的作用

细胞因子在免疫应答的产生和调节中具有重要作用,可作为免疫佐剂增强疫苗的免疫效果。近年来,重组质粒表达的细胞因子在核酸疫苗研究领域引起了人们广泛的兴趣与关注。文章简述了三类细胞因子在核酸疫苗的研究与应用,探讨了目前应用这些细胞因子类佐剂所存在的问题。     

DNA疫苗的动物试验研究及其应用前景

介绍了DNA疫苗在动物试验研究方面的现状,如:动物免疫试验中动物模型的选择、疫苗的接种途径及其选择、免疫学效果评价、安全性评价和DNA疫苗研究存在的问题,以及DNA疫苗在控制畜禽疾病方面的应用前景和发展趋势.     

A DNA vaccine against dolphin morbillivirus is immunogenic in bottlenose dolphins

The immunization of exotic species presents considerable challenges. Nevertheless, for facilities like zoos, animal parks, government facilities and non-profit conservation groups, the protection of valuable and endangered species from infectious disease is a growing concern. The rationale for immunization in these species parallels that for human and companion animals; to decrease the incidence of disease. The U.S. Navy Marine Mammal Program, in collaboration with industry and academic partners, has developed and evaluated a DNA vaccine targeting a marine viral pathogen – dolphin morbillivirus (DMV). The DMV vaccine consists of the fusion (F) and hemagglutinin (H) genes of DMV. Vaccine constructs (pVR-DMV-F and pVR-DMV-H) were evaluated for expression in vitro and then for immunogenicity in mice. Injection protocols were designed for application in Atlantic bottlenose dolphins (Tursiops truncatus) to balance vaccine effectiveness with clinical utility. Six dolphins were inoculated, four animals received both pDMV-F and pDMV-H and two animals received a mock vaccine (vector alone). All animals received an inoculation week 0, followed by two booster injections weeks 8 and 14. Vaccine-specific immune responses were documented in all four vaccinated animals. To our knowledge, this is the first report of pathogen-specific immunogenicity to a DNA vaccine in an aquatic mammal species.     

禽用DNA疫苗免疫效果增强的策略

对于人和动物的传染病来说,DNA疫苗免疫是一种很有希望的免疫方式。经过25年的发展,已经上市的核酸疫苗仍屈指可数。禽用核酸疫苗一直是禽类疫苗的研究热点,文中描述提高核酸疫苗免疫效果的方式：接种途径,疫苗剂量及首免时间,增加宿主细胞对质粒的摄入,添加免疫增强分子,优化质粒骨架和密码子,疫苗抗原的选择,异源性的首免-加强免疫策略。     

Improved immunogenicity of Newcastle disease virus inactivated vaccine following DNA vaccination using Newcastle disease virus hemagglutinin-neuraminidase and fusion protein genes

The present study describes the development of DNA vaccines using the hemagglutinin-neuraminidase (HN) and fusion (F) genes from AF2240 Newcastle disease virus strain, namely pIRES/HN, pIRES/F and pIRES-F/HN. Transient expression analysis of the constructs in Vero cells revealed the successful expression of gene inserts in vitro. Moreover, in vivo experiments showed that single vaccination with the constructed plasmid DNA (pDNA) followed by a boost with inactivated vaccine induced a significant difference in enzyme-linked immunosorbent assay antibody levels (p < 0.05) elicited by either pIRES/F, pIRES/F+ pIRES/HN or pIRES-F/HN at one week after the booster in specific pathogen free chickens when compared with the inactivated vaccine alone. Taken together, these results indicated that recombinant pDNA could be used to increase the efficacy of the inactivated vaccine immunization procedure.     

鸡柔嫩艾美球虫DNA疫苗pcDNA4．0（C）-pEtK2-IL-2对其他球虫的交叉保护力

将柔嫩艾美球虫DNA疫苗pcDNA4．0（c）-pEtK2-IL-2以50μg分别对14日龄和21日龄雏鸡进行胸部肌肉注射免疫，28日龄对各未免疫组与免疫组鸡分别口服接种柔嫩艾美球虫、巨型艾美球虫、堆形艾美球虫和毒害艾美球虫孢子化卵囊，36日龄剖杀，分析比较免疫保护效果。结果显示，柔嫩艾美球虫攻毒免疫试验组与柔嫩艾美球虫攻毒未免疫对照组比较，增重、盲肠病变记分、OPG值差异显著（P〈0．05），免疫保护效果明显，抗球虫指数（ACI）达186．50；而巨型艾美球虫、堆形艾美球虫和毒害艾关球虫攻毒免疫试验组的ACI分别为147．85、142．71和153．82，增重、盲肠病变记分、OPG值和ACI与相应的攻毒未免疫对照组比较，均有不同程度改善，免疫保护效果不明显。表明，该DNA疫苗对柔嫩艾美球虫的攻击具有完全免疫保护作用，而对巨型艾美球虫、堆形艾关球虫、毒害艾美球虫的攻击具有部分交叉保护力。     

Immune responses and expression of the virus-like particle antigen of the porcine encephalomyocarditis virus

Virus-like particles (VLPs) are particles that consist of viral capsid proteins and are structurally similar to authentic virus. To express VLPs of the porcine encephalomyocarditis virus (EMCV) and investigate their efficacy and immuno response in vivo, a plasmid (P12A3C-pCI) containing the P12A and 3C genes of the EMCV-K3 viral strain was constructed. The VLPs of EMCV-K3 were successfully assembled in 293FT cells on 3 days after transfection with P12A3C-pCI and were identified as particles of about 30-40 nm using transmission electron microscopy (TEM). In an in vivo experiment, the murine cytokines induced by VLPs of naked DNA vaccine showed that the Th1 indicators IL-2, TNF-α and GM-CSF, and the Th2 indicators IL-4 and IL-10 were increased. The immunization of mice with the P12A3C-pCI plasmid induced high levels of neutralizing antibody from 128- to 256-fold and led to a significant protection ratio (90%) after challenge with EMCV-K3 (wild-type strain). These VLPs may represent a novel vaccine strategy for the control of EMCV infection on pig farms.     

鱼类的DNA疫苗

DNA疫苗是继减毒疫苗、灭活疫苗、亚单位疫苗和重组多肽疫苗之后的又一新型疫苗，比其他疫苗高效、安全且易于大量生产。本文主要对鱼用DNA疫苗的构建、作用机制、使用方法、使用效果及安全问题等方面研究近况作一综述，以促进我国对鱼用DNA疫苗进一步的研究和应用，从而加速我国水产养殖业的发展，并为深入研究和掌握鱼类的免疫防御机制提供一些依据。     

Protection of chickens against infectious bronchitis virus with a multivalent DNA vaccine and boosting with an inactivated vaccine

The protective efficacy of DNA plasmids encoding avian infectious bronchitis virus (IBV) S1, N, or M protein was investigated in chickens. Chickens were inoculated monovalently (with plasmid pVAX1-16S1, pVAX1-16M, or pVAX1-16N alone) or multivalently (combination of the three different plasmids, pVAX1-16S1/M/N). A prime-boost immunization protocol against IBV was developed. Chickens were immunized with the multivalent DNA vaccine twice and then boosted with an inactivated vaccine once. Antibody titers of the chickens immunized with pVAX1-16S1/M/N were much higher than those of the monovalent groups (p < 0.01). A protective rate up to 90% was observed in the pVAX1-16S1/M/N group. The serum antibody titers in the prime-boost birds were significantly higher than those of the multivalent DNA vaccine group (p < 0.01) but not significantly different compared to the inactivated vaccine group at 49 days of age. Additionally, the prime-boost group also showed the highest level of IBV-specific cellular proliferation compared to the monovalent groups (p < 0.01) but no significant difference was found compared to the multivalent DNA vaccine group, and the prime-boost group completely protected from followed viral challenge.