Development of DNA vaccines for fish: vector design, intramuscular injection and antigen expression using viral haemorrhagic septicaemia virus genes as model
Development of DNA vaccines for fish: vector design, intramuscular injection and antigen expression using viral haemorrhagic septicaemia virus genes as model
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DOI：
10.1006/fsim.1997.0133
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发表时间：
1998-05-01
影响因子：
4.7
通讯作者：
Davis, HL
中科院分区：
文献类型：
--
作者：
Heppell, J;Lorenzen, N;Davis, HL
疾病控制是水产养殖业的主要关注点之一。然而，没有疫苗可用于预防许多鱼类传染病，特别是那些病毒和寄生虫来源的疾病。基于DNA的疫苗接种可以避免与传统免疫方法相关的几个问题，但对其在鱼类中的有效性知之甚少。荧光素酶和lacZ报告基因被用来表征肌肉注射虹鳟鱼和斑马鱼中质粒编码基因的表达。对于给定剂量的DNA，鱼中的荧光素酶活性高于小鼠肌肉。鱼中的酶活性在1 μ g DNA时达到峰值，并保持恒定超过12周，但它不限于注射的肌肉，因为在鳃中也检测到荧光素酶活性。注射lacZ报告基因的虹鳟鱼肌肉薄切片显示没有永久性组织损伤。为了进一步研究基于DNA的疫苗在鱼中诱导保护性免疫的能力，将病毒性出血性败血症病毒G和N基因分别克隆到表达质粒中。在转染的鱼细胞中产生的G和N蛋白似乎与天然病毒蛋白相同，因为它们被特异性单克隆抗体识别。当与对照相比时，在鱼肌肉中共注射G和荧光素酶基因导致荧光素酶活性随时间迅速降低，表明鱼对G蛋白产生细胞免疫应答，杀死转染的宿主细胞并消融CT蛋白和荧光素酶的进一步表达。最后，单独或与N构建体一起注射G构建体的幼虹鳟鱼被强烈保护免于活病毒的攻击。这些结果表明，DNA疫苗对鱼类和其他动物一样成功。(C)出版社：Academic Press Limited。
Disease control is one of the major concerns in the aquaculture industry. However, there are no vaccines available for the prevention of many piscine infectious diseases, especially those of viral and parasitic origin. DNA-based vaccination could circumvent several problems associated with traditional methods of immunization, but little is known on its efficacy in fish. The luciferase and lacZ reporter genes were used to characterize expression of plasmid-encoded genes in rainbow trout and zebra fish injected intramuscularly. For a given dose of DNA, the luciferase activity was higher in fish than in mouse muscle. The enzyme activity in fish peaked with 1 mu g of DNA and remained constant for over 12 weeks, but it was not limited to the injected muscle since luciferase activity was also detected in the gills. Thin sections of rainbow trout muscle injected with the lacZ reporter gene showed no permanent tissue damage. To further investigate the ability of DNA-based vaccines to induce protective immunity in fish, viral haemorrhagic septicaemia virus G and N genes were cloned individually into an expression plasmid. Both G and N proteins produced in transfected fish cells appeared identical to native viral proteins, as they were recognized by specific monoclonal antibodies. Coinjection of the G and luciferase genes in fish muscle resulted in a rapid decrease of the luciferase activity over time, when compared to the control, suggesting that fish raised a cellular immune response to the G protein, killing the transfected host cells and ablating further expression of CT protein and luciferase. Finally, young rainbow trout injected with the G construct, alone or together with the N construct, were strongly protected against challenge with live virus. These results suggest that DNA vaccines should be as successful for fish as they are for other animals. (C) 1998 Academic Press Limited.