PELA-OmpK微球疫苗的部分特征及其对鲫鱼的口服免疫效果  

Partial Characteristics of PELA-OmpK Microsphere Vaccine and Its Immune Effect in Crucian Carp Inoculated by Oral Route

作　　者：任燕[1] 张小江[1] 常藕琴[1] 陶家发[1] 赖迎迢[1] 石存斌[1] 吴淑勤[1] 

机构地区：[1]中国水产科学研究院珠江水产研究所,广州510380

出　　处：《中国生物制品学杂志》2011年第11期1306-1309,共4页Chinese Journal of Biologicals

基　　金：农业公益性行业科研专项(200803013);现代农业产业技术体系建设专项资金资助(nycytx-49-14);广东省科技计划项目(2007B020708004)

摘　　要：目的研究聚-DL-乳酸-聚乙二醇共聚物(DL-polylactide-co-polyethylene glycol,PELA)包裹哈维弧菌(Vibrio har-veyi,Vh)重组外膜蛋白OmpK制备的微球疫苗的部分特性及其对鲫鱼(Carassius auratus gibelio)的口服免疫效果。方法用可生物降解的合成高分子材料PELA,通过乳化溶剂挥发法包裹OmpK,制备微球疫苗,Bradford法测定蛋白浓度,计算蛋白包裹率;通过扫描电镜观察微球粒径大小及其在4℃保存不同时间的形貌。取鲫鱼随机分为3组:微球疫苗组、OmpK蛋白组和空白对照组,前两组采用疫苗或蛋白拌饲投喂方式口服免疫,投喂3 d,间隔7 d,再投喂3 d加强免疫;对照组投喂不含疫苗的饲料。加强免疫4周和8周后,经腹腔注射20 LD50的Vh EcGS020802株攻击,记录各组鱼死亡数,计算相对免疫保护率。结果制备的PELA-OmpK微球疫苗中OmpK蛋白的包裹率达78%。微球粒径小于12μm,且90%微球的粒径小于5μm。4℃保存10 d微球表面光滑、圆整;保存30 d PELA微球降解,粒径较大的微球表面出现空洞;保存60 d可见大量碎片,微球表面毛糙松散,出现多个空洞。微球疫苗加强免疫鲫鱼4周和8周后,对活菌攻击的相对免疫保护率分别为70%和48%,而口服OmpK蛋白组和空白对照组鱼全部死亡。结论用PELA包裹裸疫苗制备成微球疫苗,对抗原具有一定的保护作用,PELA用作鱼类口服疫苗的投递载体是可行的。Objective To investigate the partial characteristics of microsphere vaccine prepared by encapsulation of recombinant outer membrane protein OmpK of Vibrio harveyi （Vh） with DL-polylactide-co-polyethylene glycol （PELA） as well as its immune effect in crucian carp （Carassius auratus gibelio） inoculated by oral route. Methods Microsphere vaccine was prepared by encapsulation of OmpK with biodegradable synthetic PELA by emulsifying agent volatilization and determined for protein content by Bradford method, based on which the protein encapsulation rate was calculated. The size, as well as the morphology after storage at 4℃ for various days, of microsphere vaccine were observed by electron microscopy. Crucian carps were randomly divided into microsphere vaccine, OmpK protein and blank control groups. The crucian carps in microsphere vaccine and OmpK protein groups were fed with the forages mixed with microsphere vaccine and OmpK protein respectively for 5 d then, at an interval of 7 d, further fed for 3 d as a booster immunization, while those in blank control group were fed with vaccine-free forage. Fifty crucian carps were chosen from each group 4 and 8 weeks after booster immunization respectively and challenged with 20 LDs0 of l/h EcGS00802 strain, and the died cru- cian carps were counted, based on which the relative protective rate was calculated. Results The encapsulation rate of OmpK protein in vaccine reached 78%. The size of microsphere were less than 12 μm, of which 90% were less than 5 μm. The surfaces of microsphere after storage at 4℃ for 10 d were smooth, round and intact. However, after storage for 30 d, the PELA microsphere were degraded, and holes appeared on the surfaces of large particles. After storage for 60 d, a large quantity of debris was observed under electron microscope, while the surfaces of microspheres were rough and loose, on which several holes appeared. The relative immune protective rates of crucian carps against challenge with EcGS00802 strain were 70% and 48% 4 and

关 键 词：聚-DL-乳酸-聚乙二醇共聚物 细菌外膜蛋白质类 微球疫苗 口服免疫 

分 类 号：S941.42[农业科学—水产养殖] R392-33[农业科学—水产科学]