目的 探索氢氧化铝佐剂、N，O-CMCS纳米粒佐剂在不同免疫方式、不同免疫剂量时对柯萨奇病毒A组16型（CA16）灭活疫苗诱导小鼠体液免疫应答的影响。方法 将ICR小鼠随机分为阴性对照组，CA16灭活疫苗高、低剂量（200、40 U）组；氢氧化铝佐剂组，氢氧化铝佐剂CA16灭活疫苗高、低（200、40 U）剂量组；N，O-CMCS纳米粒佐剂组，N，O-CMCS纳米粒佐剂CA16灭活疫苗高、低（200、40 U）剂量组，共9个大组。每个大组按不同免疫方式（im、ip、id）随机分为3个小组，共计27个小组，每个小组6只小鼠（雌雄各半），注射量为200 μL/只，共免疫2次，间隔4周。分别在初次免疫后第28天、加强免疫后的第14、28天采集小鼠尾静脉血，采用微量中和试验法检测小鼠血清抗体水平，计算中和抗体几何平均滴度（GMT），中和抗体滴度≥ 1：8判定为阳性，中和抗体滴度<1：8判定为阴性，计算抗体阳转率。结果 氢氧化铝佐剂CA16灭活疫苗的抗体GMT值和中和抗体阳转率最高，无佐剂疫苗组其次，N，O-CMCS佐剂疫苗组最低；CA16灭活疫苗组的高剂量组的抗体GMT值和中和抗体阳转率均明显高于低剂量；在氢氧化铝佐剂CA16灭活疫苗组中比较中和抗体阳转率，id优于im。结论 选用氢氧化铝佐剂、CA16灭活疫苗高剂量、id免疫方式可显著增强CA16灭活疫苗诱导小鼠的体液免疫应答。

Objective To explore the effect of aluminum hydroxide adjuvant and N, O-CMCS nanoparticles adjuvant on the humoral immune response induced by CA16 inactivated vaccine in different immunization methods and doses. Method ICR mice were randomly divided into 9 groups:N, O-CMCS nanoparticles adjuvant vaccine group, aluminum hydroxide adjuvant vaccine group, non adjuvant vaccine group, aluminum hydroxide adjuvant group, N, O-CMCS nanoparticles adjuvant group and PBS control group. The vaccine was divided into high and low dose groups. Each large group was randomly divided into three groups, using the im, ip and im. 6 mice (half male and female) immunized under each immunization method. The amount of injection was 200 μL/mouse. The mice were immunized twice, with an interval of 4 weeks. The tail vein blood of mice was collected on the 28th day after primary immunization, the 14th day and the 28th day after intensive immunization, and the serum antibody level of mice was detected by micro neutralization test. The geometric mean titer (GMT) of neutralizing antibody was calculated. If the titer of neutralizing antibody was ≥ 1:8, it was positive, if the titer of neutralizing antibody was <1:8, it was negative. The positive conversion rate of antibody was calculated. Results The antibody GMT and neutralizing antibody positive rate were highest in the immunized aluminum hydroxide adjuvant vaccine, followed by the non-adjuvant vaccine group, and the lowest in the N, O-CMCS nanoparticles adjuvant vaccine group. The effect of high dose was significantly higher than that of low dose in all vaccine groups. The positive conversion rate of neutralizing antibody in id injection is better than that in intramuscular injection in the aluminum hydroxide adjuvant vaccine group. Conclusion The use of aluminum hydroxide adjuvant, high dose group, and intradermal immunization can significantly enhance the humoral immune response of mice induced by CA16 inactivated vaccine.

国家重大新药专项（2014ZX9102042001）