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1.
旨在对制备的甘露糖修饰的壳聚糖聚乳酸-羟基乙酸共聚物[poly (D,L-lactide-co-glycolide),PLGA]纳米微球作为口蹄疫病毒(foot-and-mouth disease virus,FMDV)核酸疫苗递送载体进行评价。采用西佛碱反应和元素分析制备具有一定取代度的甘露糖修饰的壳聚糖衍生物(mannose modified chitosan,MCS),然后,经双重乳化挥发法制备得到甘露糖修饰的壳聚糖PLGA纳米微球(MCS-PLGA-NPs)。采用纳米粒径仪检测MCS-PLGA-NPs粒径分布和表面电势(zeta)、扫描电镜考察其形态、琼脂糖凝胶电泳观察其对质粒的吸附和吸附质粒后抵抗核酸酶降解能力、CCK-8法检测MCS-PLGA-NPs的细胞毒性、激光共聚焦观察巨噬细胞对MCS-PLGA-NPs-质粒DNA复合物的摄取、荧光显微镜和Western blot验证MCS-PLGA-NPs加载质粒DNA在细胞中的表达。元素分析结果表明,成功制备了取代度为5%~10%的MCS。纳米粒径测定和扫描电镜结果表明,MCS-PLGA-NPs的zeta为正值、粒径分布均匀且形态规则呈球形。琼脂糖凝胶电泳结果显示,MCS-PLGA-NPs吸附质粒的能力随着其质量的增加而增强并且可以在一定程度上抵抗核酸酶降解质粒DNA。在细胞毒性试验中,不同浓度的MCS-PLGA-NPs与RAW264.7细胞共孵育24 h后,细胞存活率仍在85%以上。在细胞摄取试验中,用激光共聚焦显微镜可以明显观察到质粒DNA结合到纳米微球表面被RAW264.7细胞摄取。荧光显微镜和Western blot试验证明MCS-PLGA-NPs加载质粒DNA可以在细胞中进行表达。综上表明,本研究成功制备了MCS以及具有递送核酸疫苗能力的MCS-PLGA-NPs,为FMDV核酸疫苗的递送研究提供了新的方向和见解,也为该递送载体携带特定抗原靶向抗原递呈细胞表面甘露糖受体以及应用于动物免疫的研究奠定基础。 相似文献
2.
Vimbai Irene Machimbirike Natapol Pornputtapong Saengchan Senapin Eakapol Wangkahart Prapansak Srisapoome Pongsak Khunrae Triwit Rattanarojpong 《Journal of fish diseases》2022,45(1):1-18
Edwardsiella ictaluri infects several fish species and protection of the all the susceptible fish hosts from the pathogen using a monovalent vaccine is impossible because the species is composed of host-based genotypes that are genetic, serological and antigenic heterogenous. Here, immunoinformatic approach was employed to design a cross-immunogenic chimeric EiCh protein containing multi-epitopes. The chimeric EiCh protein is composed of 11 B-cell epitopes and 7 major histocompatibility complex class II epitopes identified from E. ictaluri immunogenic proteins previously reported. The 49.32 kDa recombinant EiCh protein was expressed in vitro in Escherichia coli BL-21 (DE3) after which inclusion bodies were successfully solubilized and refolded. Ab initio protein modelling revealed secondary and tertiary structures. Secondary structure was confirmed by circular dichroism spectroscopy. Antigenicity of the chimeric EiCh protein was exhibited by strong reactivity with serum from striped catfish and Nile tilapia experimentally infected with E. ictaluri. Furthermore, immunogenicity of the chimeric EiCh protein was investigated in vivo in Nile tilapia juveniles and it was found that the protein could strongly induce production of specific antibodies conferring agglutination activity and partially protected Nile tilapia juveniles with a relative survival percentage (RPS) of 42%. This study explored immunoinformatics as reverse vaccinology approach in vaccine design for aquaculture to manage E. ictaluri infections. 相似文献
3.
本研究旨在探索水貂肠炎病毒(mink enteritis virus,MEV)的母源抗体消长规律及其亚单位疫苗免疫效果。以免疫母貂所生仔貂为研究对象,采集21、30、45、60日龄仔貂血清,测定水貂病毒性肠炎母源抗体HI效价(MEV HI);选取25只47~52日龄健康水貂接种制备MEV基因工程亚单位疫苗,免疫前后14 d采血测定MEV HI效价;免疫后14 d进行水貂肠炎病毒攻毒临床症状观察,并测定粪便HA效价;攻毒后14 d对濒死和存活水貂安乐死,采集十二指肠、空肠和回肠进行病理组织学观察和免疫组化检测。结果显示,免疫母貂所产仔貂的MEV HI效价随着日龄的增加逐渐降低,在21日龄时较高,45日龄时少部分仔貂MEV HI效价<1:32,60日龄时大部分仔貂HI效价≤ 1:4;使用制备合格疫苗免疫后14 d对照组水貂的MEV HI效价均不高于1:4,免疫组MEV HI效价升高至1:64~1:512;攻毒保护试验表明,免疫组水貂100%抵抗水貂肠炎病毒强毒的攻击,水貂的精神、饮食、粪便等均未见异常,且攻毒后粪便HA效价为1:8~1:16;病理组织学和免疫组化检测结果表明,MEV基因工程亚单位疫苗能够很好地阻止病毒在肠黏膜上皮细胞的复制和对肠黏膜上皮细胞的损伤。因此,免疫母貂所生仔貂21日龄时体内抗体效价最高,制备的疫苗免疫50日龄左右的水貂时能够突破母源抗体干扰并产生高水平的抗体,能够抵抗水貂肠炎病毒强毒株的攻击。 相似文献
4.
5.
为定量分析贵州省临床用猪伪狂犬病(pseudorabies,PR)活疫苗中病毒含量,根据GenBank中公布的伪狂犬病病毒(pseudorabies virus,PRV)gB基因序列(登录号:M17321.1)设计1对特异性引物,建立实时荧光定量PCR方法,并对来自6个不同厂家的猪伪狂犬病活疫苗进行含毒量分析。结果显示,试验成功建立了可用于PRV检测的实时荧光定量PCR方法,标准曲线中标准品各稀释度质粒拷贝数与Ct值呈良好的线性关系,标准曲线方程为:Y=-0.97X+33.66(R2=0.999),该方法最低检测病毒含量为3.19×101拷贝/μL,敏感性高且具有良好的重复性和特异性。应用所建立的实时荧光定量PCR方法对贵州省临床用6个厂家生产的猪伪狂犬病活疫苗的病毒含量进行检测,结果显示,6种市售猪伪狂犬病活疫苗(A~F)中病毒含量分别为1.67×107、4.83×105、2.64×106、4.27×107、3.39×106和3.68×105拷贝/μL,来自不同厂家的疫苗病毒含毒量存在明显差异,最大差异可达116倍,其中来自A、D厂家的两种猪伪狂犬病活疫苗具有较高的含毒量。本试验所建立的实时荧光定量PCR方法可用于猪伪狂犬病活疫苗病毒含量的快速检测和评价,对猪伪狂犬病活疫苗的质控和临床用疫苗选择具有一定的指导意义。 相似文献
6.
7.
Protective efficacy of an H5/H7 trivalent inactivated vaccine produced from Re-11, Re-12, and H7-Re2 strains against challenge with different H5 and H7 viruses in chickens 下载免费PDF全文
ZENG Xian-ying CHEN Xiao-han MA Shu-jie WU Jiao-jiao BAO Hong-mei PAN Shu-xin LIU Yan-jing DENG Guo-hua SHI Jian-zhong CHEN Pu-cheng JIANG Yong-ping LI Yan-bing HU Jing-lei LU Tong MAO Sheng-gang GUO Xing-fu LIU Jing-li TIAN Guo-bin CHEN Hua-lan 《农业科学学报》2020,19(9):2294-2300
We developed an H5/H7 trivalent inactivated vaccine by using Re-11, Re-12, and H7-Re2 vaccine seed viruses, which were generated by reverse genetics and derived their HA genes from A/duck/Guizhou/S4184/2017(H5 N6)(DK/GZ/S4184/17)(a clade 2.3.4.4 d virus), A/chicken/Liaoning/SD007/2017(H5 N1)(CK/LN/SD007/17)(a clade 2.3.2.1 d virus), and A/chicken/Guangxi/SD098/2017(H7 N9)(CK/GX/SD098/17), respectively. The protective efficacy of this novel vaccine and that of the recently used H5/H7 bivalent inactivated vaccine against different H5 and H7 N9 viruses was evaluated in chickens. We found that the H5/H7 bivalent vaccine provided solid protection against the H7 N9 virus CK/GX/SD098/17, but only 50–60% protection against different H5 viruses. In contrast, the novel H5/H7 trivalent vaccine provided complete protection against the H5 and H7 viruses tested. Our study underscores the importance of timely updating of vaccines for avian influenza control. 相似文献
8.
余金霞 《畜牧兽医科学(电子版)》2021,(4):17-18
牛口蹄疫是传染性疾病,为研究青海省峰堆乡口蹄疫疫苗的免疫效果,于2018—2019年对该乡的7个村开展牛口蹄疫疫苗免疫效果调查,共调查牛634头,7个村总体的口蹄疫合格率为100.00%和97.56%,最低合格率为81.63%和88.63%。为更好地控制该类疾病发生流行,提升免疫效果,该文探讨口蹄疫疫苗免疫失败原因,并制定了相应的预防措施。 相似文献
9.
Feline vaccine‐associated sarcomagenesis: Is there an inflammation‐independent role for aluminium? 下载免费PDF全文
M. A. AbdelMageed P. Foltopoulou E. A. McNiel 《Veterinary and comparative oncology》2018,16(1):E130-E143
Aluminium has been found in feline vaccine‐associated sarcomas. In this study, we investigated the potential for aluminium to contribute directly to tumourigenesis. Our results indicated that an aluminium hydroxide adjuvant preparation was cytotoxic and mutagenic in human‐Chinese hamster ovary (CHO) hybrid cells in vitro. Moreover, CHO cells deficient in DNA double strand break (DSB), but not single‐strand break (SSB), repair, were particularly sensitive to aluminium exposure compared with repair proficient cells, suggesting that aluminium is associated with DSBs. In contrast to CHO cells, primary feline skin fibroblasts were resistant to the cytotoxic effects of aluminium compounds and exposure to an aluminium chloride salt promoted cell growth and cell cycle progression at concentrations much less than those measured in particular feline rabies vaccines. These findings suggest that aluminium exposure may contribute, theoretically, to both initiation and promotion of tumours in the absence of an inflammatory response. 相似文献
10.
Goatpox (GTP), sheeppox (SPP) and lumpy skin disease (LSD) are three severe diseases of goat, sheep and cattle. Their typical clinical symptoms are characterized by vesicles, papules, nodules, pustules and scabs on animal skins. The GTP, SPP and LSD are caused by goatpox virus (GTPV), sheeppox virus (SPPV) and lumpy skin disease virus (LSDV), respectively, all of which belong to the genus Capripoxvirus in the family Poxviridae. Several capripoxvirus (CaPV) isolates have been virulently attenuated through serial passaging in vitro for production of live vaccines. CaPV-based vector systems have been broadly used to construct recombinant vaccines for delivering foreign antigens, many of which have been demonstrated to induce effective immune protections. Homologous recombination is the most commonly used method for constructing recombinant CaPVs. Here, we described a methodology for generation of recombinant CaPVs by the homologous recombination, and further reviewed CaPV-vectored vaccines for delivering foreign antigens. 相似文献