Development of oral DNA vaccine based on chitosan nanoparticles for the immunization against reddish body iridovirus in turbots (Scophthalmus maximus)
Development of oral DNA vaccine based on chitosan nanoparticles for the immunization against reddish body iridovirus in turbots (Scophthalmus maximus)
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DOI：
10.1016/j.aquaculture.2015.11.013
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发表时间：
2016-02
期刊：
影响因子：
4.5
通讯作者：
Fengrong Zheng;Hongzhan Liu;Xiu-qin Sun;Yongqiang Zhang;Baiyu Zhang;Zhaojun Teng;Hou Yongjiang;Bo Wang
中科院分区：
文献类型：
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作者：
Fengrong Zheng;Hongzhan Liu;Xiu-qin Sun;Yongqiang Zhang;Baiyu Zhang;Zhaojun Teng;Hou Yongjiang;Bo Wang
大菱鲆红体病虹彩病毒（TRBIV）是一种引起大菱鲆病毒性红体综合征（VRBS）的鱼类虹彩病毒。病毒的感染严重阻碍了大菱鲆养殖业的发展。因此，研制预防性疫苗至关重要。在前期研究中，我们构建了重组质粒pEGFP-N2-TRBIV-MCP，并对其免疫效果进行了评价。本研究以大菱鲆（Scophthalmus maximus）为试验动物，研究了TRBIV纳米颗粒口服DNA疫苗的免疫效果。采用离子凝胶法制备了包裹质粒DNA的壳聚糖纳米粒，并通过透射电镜和MalvinZS分析仪对壳聚糖纳米粒的性质进行了分析。结果表明，壳聚糖纳米粒为球形，粒径为145.5 ± 1.5 nm，zeta电位为24.3 ± 0.5。将pDNA-CS-NPs体外转染大菱鲆TK细胞系，RT-PCR和激光共聚焦显微镜检测pDNA-CS-NPs的瞬时表达。经口给药后，在大菱鲆的不同组织中均可检测到抗原基因的表达。maximus）。pDNA-CS-NPs能明显诱导小鼠产生特异性免疫应答，并能显著上调MHCIα、IIα、IFN-1、IFN-g、Mx-1、IL-8和Mx的表达。经口免疫的大菱鲆免疫保护力较强，相对存活率为68%，免疫后90 d内可产生可检测水平的抗TRBIV抗体。这些结果表明，pDNA-CS-NPs是一种很有前途的口服疫苗载体。该方法可以推广到其他口服疫苗的开发，并应用于其他鱼类抗TRBIV。相关声明在这项工作中，开发了一种口服的基于纳米颗粒的DNA疫苗，以评估免疫效果在大菱鲆（S。maximus）。采用离子凝胶法制备了包裹质粒DNA的壳聚糖纳米粒，并通过透射电镜和MalvinZS分析仪对壳聚糖纳米粒的性质进行了分析。我希望这篇论文适合水产养殖。
Turbot reddish body iridovirus (TRBIV), a piscine iridovirus, causes viral reddish body syndrome (VRBS) in turbot and is highly lethal to the fish. Infection of the virus hampers the development of turbot aquaculture. Hence, it is important to develop a preventive vaccine. In our previous study, we constructed and evaluated the immunization efficacy of the recombinant plasmid, pEGFP-N2-TRBIV-MCP. In the present study, an oral nanoparticle-based DNA vaccine against TRBIV was developed to evaluate the immunization effectiveness in turbots (Scophthalmus maximus) through oral administration. We prepared chitosan nanoparticles enwrapping plasmid DNA using ionic gelation process, and the characteristics of chitosan nanoparticles were analyzed by transmission electron microscope and Malvin ZS analyzer. The results revealed that the chitosan nanoparticle plasmids encoding DNA (pDNA-CS-NPs) were spherical shape with the particle size and zeta potential of 145.5 ± 1.5 nm and 24.3 ± 0.5, respectively. The loading rate and encapsulation efficiency were 92.8 ± 1.38% and 63.7 ± 0.89%, respectively.In vitrotransfection, pDNA-CS-NPs was transferred into eukaryotic cells, turbot cell TK line, and transient expression of pDNA-CS-NPs was detected by RT-PCR and laser scanning confocal microscope. After oral administration, the expression of the antigen gene could be detected in different tissues of turbots (S. maximus) at 20 and 40 days post-vaccination. The pDNA-CS-NPs clearly induced an innate and specific immune response, and significantly up-regulated the expression of MHCIα, IIα, IFN-1, IFN-g, Mx-1, IL8 and Mx. In addition, strong protection with relative survival rate of 68% was recorded in the orally-vaccinated turbots, which could produce the detectable level of anti-TRBIV antibody during 90 days. These data in this study suggested that pDNA-CS-NPs were promising carriers for plasmid DNA vaccine through oral vaccination. This method could be extended to the development of other oral vaccines and applied in other fish species against TRBIV.Statement of relevanceIn this work, an oral nanoparticle-based DNA vaccine against TRBIV was developed to evaluate the immunization effectiveness in turbots (S. maximus) through oral administration. We prepared chitosan nanoparticles enwrapping plasmid DNA using ionic gelation process, and the characteristics of chitosan nanoparticles were analyzed by transmission electron microscope and Malvin ZS analyzer. I hope this paper is suitable for aquaculture.