狂犬疫苗脂质体冻干粉制备及免疫原性研究  

Preparation and immunogenicity of lyophilized rabies virus liposome vaccine

作　　者：叶红艳[1] 徐菀佚 张益丽 马波 鲁卫东[1] 

机构地区：[1]昆明医科大学药学院/云南省天然药物药理重点实验室,云南昆明650500 [2]江苏沃森生物技术有限公司,江苏泰州225300

出　　处：《扬州大学学报（农业与生命科学版）》2016年第3期11-14,共4页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：江苏省科技型企业技术创新资金项目(BC2011194)

摘　　要：采用薄膜分散联合冻融冻干法制备狂犬疫苗脂质体冻干粉,以包封率为主要考察指标,通过正交试验确定制备的最佳工艺,并考察其粒度分布、包封率及免疫原性;用狂犬疫苗脂质体和疫苗原液分别免疫小鼠,快速荧光灶抑制试验法检测中和抗体,体外试验法检测淋巴细胞增殖能力,流式细胞仪检测淋巴细胞表面标记。结果表明:通过工艺优化所制备的狂犬疫苗脂质体冻干粉复溶后呈圆形或椭圆形,粒度分布均匀,平均粒径约为4.36μm,包封率为82.96%;中和抗体产生情况在前14d疫苗脂质体组明显高于疫苗原液组,后28d2组产生的中和抗体情况相似;狂犬疫苗脂质体组的淋巴细胞增殖指数和小鼠CD4+/CD8+比例与疫苗原液组相比,差异均显著。证明狂犬疫苗脂质体能够较早地产生中和抗体以提高对机体的保护效力,其不仅可增强体液免疫且可增强细胞免疫。Rabies vaccine lyophilized liposome was prepared based on the method of film-dispersion and freeze-thawing method.Orthogonal design method was adopted to optimize the formulation by using encapsulation efficiency as the evaluation index.The particle distribution,encapsulation efficiency and immunogenicity of the resulted product were investigated.Mice were immunized with rabies vaccine lyophilized liposome and adjuvant-free rabies vaccine respectively,then the neutralizing antibody was determined by RFFIT,the proliferation of lymphocyte was carried on by in vitro test,and the surface marker of lymphocytes was obtained by flow cytometry.The results showed that,the prepared rabies vaccine liposome were round or elliptic,and the particles were evenly distributed at a mean size of 4.36μm with the encapsulation efficiency of 82.96%.The neutralizing antibody level of mice immunized with liposome vaccine were significantly higher than those with adjuvant-free rabies vaccine at the first 14 d.After that till 28 d,the neutralizing antibody level from two groups of mice were similar.Compared with adjuvant-free vaccine immunized mice group,the lymphocyte proliferation index and the proportion of CD4＋/CD8＋from rabies vaccine liposome immunized mice group were much higher.The data proved that rabies vaccine lyophilized liposome was capable to improve the protection effect of body by producing the neutralizing antibody in early days.Both the humoral and the cellular immunity could be strengthened by the resulted product.

关 键 词：Vero细胞狂犬疫苗 脂质体 体液免疫 细胞免疫 

分 类 号：R512.99[医药卫生—内科学] S852.65[医药卫生—临床医学]