一种口蹄疫灭活疫苗水包油复乳佐剂的制备及其皮内免疫效果评价  

Preparation of an oil-in-water emulsion adjuvant for inactivated foot-and-mouth disease vaccine and its intradermal immune effect evaluation

作　　者：许泽玉 尹文竹 周宇杰 邓碧华[2,3] 卢宇 杨平[1] XU Zeyu;YIN Wenzhu;ZHOU Yujie;DENG Bihua;LU Yu;YANG Ping(College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China;Institute of Veterinary Immunology&Engineering,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;Guotai(Taizhou)Center of Technology Innovation for Veterinary Biologicals,Taizhou 225300,China)

机构地区：[1]南京农业大学动物医学院,江苏南京210095 [2]江苏省农业科学院动物免疫工程研究所,江苏南京210014 [3]兽用生物制品(泰州)国泰技术创新中心,江苏泰州225300

出　　处：《中国兽医科学》2024年第6期766-776,共11页Chinese Veterinary Science

基　　金：江苏省农业科技自主创新资金项目(CX(22)2018);江苏省重点研发计划项目(BE2020336)。

摘　　要：为了提高口蹄疫灭活疫苗的免疫效果,本研究对佐剂选择和免疫途径进行了疫苗的安全性和免疫效果评价,最终筛选出油酸乙酯、聚氧乙烯月桂醇及PEG400以2∶5∶3的质量比例混合形成复乳佐剂(简称OEPP),配伍口蹄疫灭活抗原,通过皮内免疫小鼠,检测其安全性并探究其免疫效果。结果显示,OEPP可以在4~60℃下稳定存放30 d;体外溶血试验证明,OEPP不会造成红细胞破裂,且红细胞形态良好。进一步通过组织病理学分析OEPP不会诱发重要生理器官的病变。体内外安全测试均证实OEPP具有较好的生物相容性,可以用于体内注射使用;皮内免疫OEPP与商品化ISA206后血清中FMDV-Ig G抗体效价无显著性差异,但显著高于肌肉免疫;此外,与肌肉免疫相比,发现皮内免疫可诱导注射部位皮肤组织募集更多的淋巴细胞、炎性细胞。结果表明,复乳佐剂OEPP经皮内免疫可以诱导局部炎性反应,诱导更多抗原递呈细胞聚集,促进了免疫应答效力,为后续本体动物的无针皮内免疫试验提供了试验数据。In this study,adjuvants and immune pathways have been investigated to assess the safety and immunological efficacy of the inactivated foot-and-mouth disease(FMD)vaccine,aiming to enhance its effectiveness.The oil-in-water(O/W)emulsion adjuvant(short as OEPP)was composed of ethyl oleate,polyoxyethylene lauryl alcohol,and PEG400 at a mass ratio of 2∶5∶3 and combined with inactivated foot-and-mouth disease virus(FMDV)by intradermal injection to explore its biosafety and immune effect.From the findings,OEPP could be kept steadily at 4℃to 60℃for 30 days without stratification.According to the results of the hemolysis test in vitro,the red blood cells treated with OEPP did not burst and were in good condition.Additionally,H&E examination demonstrated that OEPP did not trigger pathological changes in essential physiological organs.All safety tests in vivo and in vitro confirmed that OEPP has high biocompatibility and may be utilized for in vivo injection.Importantly,there was no significant difference in the level of FMDV-Ig G antibody between OEPP and commercial ISA206,but it was significantly higher than intramuscular immunity.Also,intradermal immunity could recruit more lymphocytes and inflammatory cells in the skin tissue surrounding the injection site than intramuscular injection.OEPP can induce a local inflammatory reaction through the intradermal immunological pathway,thereby recruiting more antigen-presenting cells(APCs)and promoting immune response-ability.This may provide experimental data for subsequent needle-free intradermal immunization tests in vivo.

关 键 词：口蹄疫 复乳佐剂 皮内注射 疫苗 

分 类 号：S859.797[农业科学—临床兽医学]