犬细小病毒VP2蛋白在家蚕/昆虫细胞中表达  

Expression of the VP2 gene of canine parvovirus in BmN cells/silkworm pupae

作　　者：胡桂秋[1,2,3] 郭鹤[1,2] 梁红茹[1,2] 冯昊[1,2] 王化磊[1,2] 郑学星[1,2] 赵永坤[1,2] 赵丽丽[1,2] 杨松涛[1,2] 夏咸柱[1,2] 

机构地区：[1]军事医学科学院军事兽医研究所,吉林长春130122 [2]吉林省人兽共患病预防与控制重点实验室,吉林长春130062 [3]吉林农业大学,吉林长春130118

出　　处：《中国病原生物学杂志》2013年第7期577-580,共4页Journal of Pathogen Biology

基　　金：国家公益性行业(农业)科研专项(No.201303042)

摘　　要：目的犬细小病毒(canine parvovirus,CPV)能够引起犬急性出血性肠炎,在幼犬中的死亡率很高,为我国养犬业和经济动物养殖业带来巨大的经济损失。因而,寻找一种新型、安全、高效的疫苗为细小病毒性肠炎的防控具有重要意义。方法本研究利用Bac-to-Bac表达系统构建了表达CPV VP2蛋白的重组杆状病毒rBV-D-VP2。该重组病毒感染昆虫细胞和家蚕后,表达CPV VP2蛋白。体外表达的VP2蛋白自动装配成病毒样粒子。随后用蚕蛹中大量组装成的病毒样粒子分别通过口服和肌注两种途径免疫豚鼠。结果经电镜检测,观察到直径为25nm大小的球形病毒样粒子,表明重组杆状病毒表达的VP2能形成病毒样粒子,且在大小、形态上与天然病毒相似。间接免疫荧光检测表明成功构建了表达VP2的病毒样粒子。口服和肌注两种途径免疫豚鼠,均产生了血凝抑制效价。结论本研究为CPV新型亚单位疫苗的研究提供了依据。Objective Canine parvovirus causes acute hemorrhagic gastroenteritis in pups, leading to a high mortality rate. This results in vast economic losses for animal breeders. A new safe and effective vaccine to prevent canine parvovir- us must be found. Methods A recombinant baculovirus named rBV-D-VP2 was constructed using a Bac-to-Bac ex- pression system. The VP2 protein of canine parvovirus was expressed in insect cells and silkworm pupae and self-assem- bled into virus-like particles （VLPs）. Silkworm homogenates containing CPV-VLPs were then administered to guinea pigs orally and intramuscularly. Results Electron microscopy showed that the VP2 protein assembled into spherical particles with a diameter of 25 nm~ these particles were similar to native CPV in terms of both size and morphology. Indi- rect immunofluorescence showed that the VP2 protein was successfully expressed in BmN cells. Oral and intramuscular delivery induced a humoral immune response in guinea pigs. Conclusion This study lays the foundation for the pro- duction of new subunit vaccines against CPV in the future.

关 键 词：犬细小病毒 VP2 病毒样粒子 免疫原 

分 类 号：S852.65[农业科学—基础兽医学]