Rajesh Kumar S, Ishaq Ahamed V P, Sarathi M, Nazeer Basha A, Sahul Hameed A S
Aquaculture Biotechnology Division, Department of Zoology, C. Abdul Hakeem College, Melvisharam-632 509, Vellore District, Tamil Nadu, India.
Fish Shellfish Immunol. 2008 Apr;24(4):467-78. doi: 10.1016/j.fsi.2008.01.004. Epub 2008 Jan 11.
White spot disease is an important viral disease caused by white spot syndrome virus (WSSV) and is responsible for huge economic losses in the shrimp culture industry worldwide. The VP28 gene encoding the most dominant envelope protein of WSSV was used to construct a DNA vaccine. The VP28 gene was cloned in the eukaryotic expression vector pcDNA3.1 and the construct was named as pVP28. The protective efficiency of pVP28 against WSSV was evaluated in Penaeus monodon by intramuscular challenge. In vitro expression of VP28 gene was confirmed in sea bass kidney cell line (SISK) by fluorescence microscopy before administering to shrimp. The distribution of injected pVP28 in different tissues of shrimp was studied and the results revealed the presence of pVP28 in gill, head soft tissue, abdominal muscle, hemolymph, pleopods, hepatopancreas and gut. RT-PCR and fluorescence microscopy analyses showed the expression of pVP28 in all these tissues examined. The results of vaccination trials showed a significantly higher survival rate in shrimp vaccinated with pVP28 (56.6-90%) when compared to control groups (100% mortality). The immunological parameters analyzed in the vaccinated and control groups revealed that the vaccinated shrimp showed significantly high level of prophenoloxidase and superoxide dismutase (SOD) when compared to the control groups. The high levels of prophenoloxidase and superoxide dismutase (SOD) might be responsible for developing resistance against WSSV in DNA vaccinated shrimp.
白斑病是由白斑综合征病毒(WSSV)引起的一种重要病毒性疾病,在全球对虾养殖业造成了巨大经济损失。编码WSSV最主要包膜蛋白的VP28基因被用于构建DNA疫苗。VP28基因被克隆到真核表达载体pcDNA3.1中,构建体被命名为pVP28。通过肌肉注射攻毒在斑节对虾中评估pVP28对WSSV的保护效率。在将其注射给对虾之前,通过荧光显微镜在鲈鱼肾细胞系(SISK)中证实了VP28基因的体外表达。研究了注射的pVP28在对虾不同组织中的分布,结果显示在鳃、头部软组织、腹肌、血淋巴、腹足、肝胰腺和肠道中存在pVP28。RT-PCR和荧光显微镜分析表明pVP28在所检测的所有这些组织中均有表达。疫苗接种试验结果显示,与对照组(100%死亡率)相比,用pVP28接种的对虾存活率显著更高(56.6 - 90%)。对接种组和对照组分析的免疫参数显示,与对照组相比,接种疫苗的对虾显示出显著高水平的前酚氧化酶和超氧化物歧化酶(SOD)。高水平的前酚氧化酶和超氧化物歧化酶(SOD)可能是DNA疫苗接种的对虾对WSSV产生抗性的原因。