Al(OH)_3佐剂对不同抗原含量肠道病毒71型灭活疫苗免疫效果的影响  

Influence of aluminium hydroxide adjuvant on immune effect of inactivated enterovirus 71 vaccine containing various concentrations of antigen

作　　者：李华[1] 姜广菊 杨婷[1] 何晓娟[1] 岳磊[1] 谢天宏[1] 杨水芝[1] 朱凡丽 龙润乡[1] 杨蓉[1] 罗芳宇 谢忠平[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省重大传染病疫苗工程技术研究中心云南省重大传染病疫苗研发重点实验室,云南昆明650118

出　　处：《中国生物制品学杂志》2015年第2期105-109,114,共6页Chinese Journal of Biologicals

基　　金：云南省重点新产品研究项目"国家III类医疗器械手足口病EV71及Cox A16病毒抗原体外诊断试剂的临床试验研究"(2012BC006)

摘　　要：目的分析Al(OH)3佐剂对不同抗原含量肠道病毒71型(enterovirus 71,EV71)灭活疫苗免疫效果的影响。方法将ICR小鼠随机分为9组,每组8只:常规剂量(160 EU/0.1 ml EV71)、1/2剂量(80 EU/0.1 ml EV71)、1/5剂量(32 EU/0.1 ml EV71)无佐剂及佐剂[Al(OH)31.0 mg/ml]疫苗组,并设佐剂、生理盐水及空白对照组,分别于0、28 d经肌肉免疫小鼠,分别于初免及加强免疫后28 d,经尾静脉采血,分离血清,采用体外微量中和试验法检测小鼠血清中和抗体效价,流式细胞仪检测IL-2、IL-4、IL-6、IL-17A、IL-10、IFNγ及TNF水平。结果初免后28 d,1/2剂量疫苗组中佐剂组小鼠血清抗体阳性率明显高于无佐剂疫苗组,差异有统计学意义(P<0.05),其余各组间差异无统计学意义(P>0.05);加强免疫后28 d,常规剂量疫苗组中,佐剂及无佐剂组小鼠血清抗体阳性率均为100%,抗体GMT分别为1∶152.2和1∶139.6,1/2剂量疫苗组中,佐剂组小鼠血清中抗体GMT与无佐剂组相比,差异有统计学意义(P<0.05),其余组间抗体阳性率及抗体GMT差异均无统计学意义(P>0.05)。常规剂量、1/2剂量、1/5剂量疫苗组,均诱导产生IL-6、IL-10、IFNγ及TNF 4种细胞因子,与佐剂对照组和生理盐水对照组相比,差异有统计学意义(P﹤0.05),未检测到IL-2、IL-4、IL-17A。结论 EV71灭活疫苗可诱导小鼠产生IL-6、IL-10、IFNγ、TNF 4种细胞因子及特异性的抗EV71抗体,加入佐剂后免疫效果更好。Objective To analyze the influence of aluminium hydroxide adjuvant on immune effect of inactivated enterovirus 71（EV71） vaccine containing various concentrations of antigen. Methods ICR mice were randomly divided into nine groups,8 for each. The mice in six test groups were immunized with adjuvant-free and adjuvant-containing（1. 0 mg / ml aluminium hydroxide）inactivated EV71 vaccine at routine dosage（160 EU / 0. 1 ml EV71）,1 / 2 dosage（80 EU / 0. 1 ml EV71）and 1 / 5 dosage（32 EU / 0. 1 ml EV71）respectively,while those in three control groups were immunized with aluminium hydroxide adjuvant and physiological saline and unimmunized（blank）respectively. Mice were immunized i. m. twice on days 0 and 28,of which the serum samples were collected 28 d after primary and booster immunizations respectively,and determined for neutralizing antibody titer by microneutralization test in vitro,and for IL-2,IL-4,IL-6,IL-17 A,IL-10,IFNγ and TNF levels by flow cytometry. Results The serum antibody positive rate was significantly higher in mice 28 d after primary immunization with adjuvant-containing vaccine at 1 / 2 dosage than in those with adjuvant-free vaccine（P 〈 0. 05）,while showed no significant difference in other groups（P 〉 0. 05）. Both the serum antibody positive rates of mice 28 d after booster immunization at routine dosage were 100%,while the GMTs were1 ∶ 152. 2 and 1 ∶ 139. 6 respectively. The GMTs of antibodies induced by adjuvant-containing vaccines at 1 / 2 dosage showed significant difference with those by adjuvant-free vaccines（P 〈 0. 05）. However,no significant differences were observed in positive rates and GMTs of antibodies in other groups（P 〉 0. 05）. The vaccines at routine,1 / 2 and 1 / 5dosages induced Il-6,IL-10,IFNγ and TNF,which showed significant difference with those in adjuvant and physiological saline control groups（P 〈 0. 05）. However,no IL-2,IL-4 or IL-17 A was detected. Conclusion Inactivated EV71 vaccine induced specific anti-EV71 antibo

关 键 词：肠道病毒71型 疫苗 佐剂 免疫 

分 类 号：R373.2[医药卫生—病原生物学] R392-33[医药卫生—基础医学]