鳗鲡病原性嗜水气单胞菌与爱德华氏菌二联表达外膜蛋白的免疫原性

发布时间：2018-05-09 00:19

本文选题：二联外膜蛋白 + 嗜水气单细胞菌　；参考：《中国水产科学》2015年01期

【摘要】：采用鳗鲡源嗜水气单胞菌(Aeromonas hydrophila)和迟缓爱德华氏菌(Edwardsiella tarda)外膜蛋白基因二联表达产物免疫日本鳗鲡(Anguilla japonica),检测其对日本鳗鲡免疫功能的影响及其攻毒免疫保护力。将150尾日本鳗鲡平均分为PBS、细菌免疫和外膜蛋白免疫3个组,3组鳗鲡分别以PBS(0.01 mol/L,p H7.4)、嗜水气单胞菌与迟缓爱德华氏菌二联灭活苗(5.0×108 CFU/m L)、嗜水气单胞菌与迟缓爱德华氏菌外膜蛋白二联表达产物(500μg/m L)腹腔注射0.2 m L。于免疫后14、21和28 d麻醉鳗鲡采血并分离抗凝血。测定3个时间点鳗鲡血浆中特异性抗体效价和同时期鳗鲡血浆、体表黏液、肝和肾组织匀浆液中的溶菌酶含量,同时检测3个时间点鳗鲡全血细胞的转化水平。免疫后28 d,嗜水气单胞菌和迟缓爱德华氏菌分别腹腔注射感染3组鳗鲡并测定其相对免疫保护率。结果表明,免疫后14 d和28 d,灭活菌和外膜蛋白免疫组的抗体水平均极显著高于PBS组(P0.01)。溶菌酶检测结果表明,不同处理组不同时间段血清、黏液和肝肾组织悬液的溶菌酶含量存在显著(P0.05)或极显著(P0.01)差异。免疫后14 d灭活菌组全血细胞转化水平显著高于PBS和外膜蛋白组(P0.05),而21 d两个免疫组则均显著低于PBS组(P0.05)。活菌感染结果表明,灭活菌和外膜蛋白免疫后28 d对两株病原菌的攻毒相对免疫保护率均比PBS组提高了50%(P0.05)。本研究结果表明鳗鲡源嗜水气单胞菌和迟缓爱德华氏菌外膜蛋白二联表达产物免疫日本鳗鲡后可提高鳗鲡的免疫功能及其对这两株菌的抵抗力,从而可能应用于鳗鲡基因工程疫苗的研发。
[Abstract]:Anguilla japonica was immunized with the double expression products of outer membrane protein gene of Aeromonas hydrophila and Edwardsiella tarda. 150 Japanese eels (Anguilla japonica) were divided into three groups on average: PBS, bacterial immunization and outer membrane protein immunization. The eels were treated with PBS(0.01 mol-lpH7.4, Aeromonas hydrophila and Edwards tardy inactivated vaccine 5.0 脳 10 ~ 8 CFU/m / L, Aeromonas hydrophila and Edward tardy respectively. The diplex expression product of outer membrane protein was injected intraperitoneally with 0.2 mL of 500 渭 g 路mL ~ (-1) 路L ~ (-1) 路L ~ (-1). The blood samples were collected and anticoagulant blood was isolated from anaesthetized Anguilla japonica on day 14, 21 and 28 after immunization. The specific antibody titers in plasma of Anguilla japonica and the lysozyme contents in plasma, body surface mucus, liver and kidney homogenate of Anguilla japonica were measured at three time points, and the transformation level of whole blood cells of Anguilla japonica at three time points was also measured. On the 28th day after immunization, Aeromonas hydrophila and Edward's retardation were injected intraperitoneally into three groups of eels, and their relative immune protection rates were measured. The results showed that the antibody levels of inactivated bacteria and outer membrane protein immunized group were significantly higher than that of PBS group on the 14th and 28th day after immunization. The results of lysozyme detection showed that there were significant differences in lysozyme content in serum, mucus and liver and kidney suspension between different treatment groups (P 0.05) or extremely significant (P 0.01). On the 14th day after immunization, the whole blood cell transformation level in inactivated bacteria group was significantly higher than that in PBS group and outer membrane protein group (P0.05A), but on the 21st day, it was significantly lower in both groups than that in PBS group. The results of live bacteria infection showed that the relative immunity protection rate of inactivated bacteria and outer membrane protein against the two pathogens 28 days after immunization was higher than that of PBS group (P 0.05). The results showed that the double expression products of Aeromonas hydrophila and Edwardsiella tarda could improve the immune function of Anguilla japonica and their resistance to the two strains of Anguilla japonica. Therefore, it may be used in the development of genetic engineering vaccine of Anguilla japonica.
【作者单位】：集美大学水产学院鳗鲡现代产业技术教育部工程研究中心;
【基金】：国家自然科学基金项目(31001136)
【分类号】：S943

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