流感H3N2疫苗纳米乳佐剂的优化及安全性  

Optimization and safety of nano-emulsion adjuvant of influenza H3N2 vaccine

作　　者：赵兰华[1] 刘洪卿[1] 钏鸿云 段男 杨姝[1] 杨磊[1] 戴宗祥[1] 李映波[1] 

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所,云南昆明650118

出　　处：《中国生物制品学杂志》2014年第6期775-779,共5页Chinese Journal of Biologicals

基　　金：十一五国家高技术研究发展计划(863)基金资助项目(2006AA02Z409);云南省科技计划项目(计划类别:联合支持国家项目)(2008GA007)

摘　　要：目的 优化纳米乳新配方,并从载水量及载苗方式探讨影响佐剂效果的因素。方法 将ICR小鼠随机分为7组,每组5只,即A1、A2、A3、A4纳米乳佐剂组[A1（载水量66.7%）、A2（载水量80.0%）、A3（载水量85.7%）为成乳后加入疫苗吸附,A4（载水量80.0%）为成乳前加入疫苗包裹]：分别加入等量H3N2裂解疫苗[血凝素（hemagglutinin,HA）1.8μg];空白对照组：生理盐水;单独疫苗组：HA 1.8μg;铝佐剂组：A（lOH）30.2 mg,HA 1.8μg。均经小鼠大腿腓肠肌注射0.2 ml,0、21 d免疫,于初免后第1周开始,每周经尾动脉采血,分离血清,检测血凝抑制（haemagg-lutination inhibition,HI）效价,比较各组的免疫应答水平。以初免后1周内小鼠体重变化及最大给药量试验分析纳米乳的最大耐受量（maximal tolerance dose,MTD）。采用马尔文粒度仪检测不同载水量纳米乳粒径大小及分散系数。伪三元相图法及静置观察检测稳定性。结果 小鼠初免后第5周,单独疫苗组小鼠HI滴度与A1、A2、A3纳米乳佐剂组相比,差异有统计学意义（P〉0.05）,与A4纳米乳佐剂组相比,差异无统计学意义（P〈0.05）;A3纳米乳佐剂组于初免后第5周,HI滴度达到峰值,〉为单独疫苗组的4倍,也明显高于铝佐剂对照组和A1、A2纳米乳佐剂组（P均〉0.05）。纳米乳搭载或包裹疫苗对小鼠生长无明显影响,MTD〈25 ml/kg。纳米乳平均粒径分布均为10～100 nm左右,纳米乳A1、A2、A3粒径大小无明显差异,均匀度较好。载水范围大的纳米乳可搭载足量抗原,稳定性好。结论 载水量85.7%、以吸附方式载苗的纳米乳佐剂效果最佳,且具有良好的安全性。Objective To optimize the formula of nano-emulsion as an adjuvant of influenza H3N2 vaccine and investigate the influencing factors of adjuvant effect using water-loading and mode of vaccine loading as indexes. Methods ICR mice were divided into seven groups. The mice in A1, A2, A3 and A4 groups were injected i. m. with the mixture of nano-emulsion adjuvant and influenza H3N2 split vaccine containing 1.8 μg hernagglutinin （HA）, in a vol- ume of 0. 2 ml, on days 0 and 21. The vaccines in A1, A2 and A3 groups were added after emulsion, while that in A4 group before emulsion. The water-loadings in A1, A2, A3 and A4 groups were 66. 7%, 80. 0%, 85. 7% and 80. 0% respectively. However, the mice in blank control group were injected with physiological saline, while those in vaccine control group with 1.8 μg HA, and those in aluminum adjuvant control group with 0. 2 mg aluminum hydroxide ＋ 1.8 μg HA. Blood samples were collected every week starting from week 1 after the first injection and determined for haemagglutination inhibition （HI） antibody titer, based on which the immune response levels in various groups were com- pared. The maximal tolerance dose （MTD） of nano-emulsion was analyzed by the change of bodyweights of mice within one week after the first injection and maximal dose test. The sizes and dispersion coefficients of nano-emulsion particles with various water-loadings were analyzed by Melvin granulometer. The stability was evaluated by pseudo ternary phase diagram method. Results The HI titer of mice in vaccine control group at week 5 after the first injection showed significant difference with those in A1, A2 and A3 groups （P 〈 0. 05）, while showed no significant difference with that in A4 group （P 〉 0. 05 ）. The HI titer in A3 group reached the maximum at week 5 after the first injection, whichwas about 4 times of that in vaccine control group, and were significantly higher than those in aluminum adjuvant control, A1 and A2 groups （each P 〈 0. 05 ）. Loading or embedding o

关 键 词：纳米乳 佐剂 流感疫苗 优化 安全性 

分 类 号：R373.13[医药卫生—病原生物学] R392-33[医药卫生—基础医学]