目的 探索氫氧化鋁佐劑、N，O-CMCS納米粒佐劑在不同免疫方式、不同免疫劑量時(shí)對柯薩奇病毒A組16型（CA16）滅活疫苗誘導(dǎo)小鼠體液免疫應(yīng)答的影響。方法 將ICR小鼠隨機(jī)分為陰性對照組，CA16滅活疫苗高、低劑量（200、40 U）組；氫氧化鋁佐劑組，氫氧化鋁佐劑CA16滅活疫苗高、低（200、40 U）劑量組；N，O-CMCS納米粒佐劑組，N，O-CMCS納米粒佐劑CA16滅活疫苗高、低（200、40 U）劑量組，共9個(gè)大組。每個(gè)大組按不同免疫方式（im、ip、id）隨機(jī)分為3個(gè)小組，共計(jì)27個(gè)小組，每個(gè)小組6只小鼠（雌雄各半），注射量為200 μL/只，共免疫2次，間隔4周。分別在初次免疫后第28天、加強(qiáng)免疫后的第14、28天采集小鼠尾靜脈血，采用微量中和試驗(yàn)法檢測小鼠血清抗體水平，計(jì)算中和抗體幾何平均滴度（GMT），中和抗體滴度≥ 1：8判定為陽性，中和抗體滴度<1：8判定為陰性，計(jì)算抗體陽轉(zhuǎn)率。結(jié)果 氫氧化鋁佐劑CA16滅活疫苗的抗體GMT值和中和抗體陽轉(zhuǎn)率最高，無佐劑疫苗組其次，N，O-CMCS佐劑疫苗組最低；CA16滅活疫苗組的高劑量組的抗體GMT值和中和抗體陽轉(zhuǎn)率均明顯高于低劑量；在氫氧化鋁佐劑CA16滅活疫苗組中比較中和抗體陽轉(zhuǎn)率，id優(yōu)于im。結(jié)論 選用氫氧化鋁佐劑、CA16滅活疫苗高劑量、id免疫方式可顯著增強(qiáng)CA16滅活疫苗誘導(dǎo)小鼠的體液免疫應(yīng)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.

國家重大新藥專項(xiàng)（2014ZX9102042001）