氢氧化铝佐剂对肠道病毒71型灭活疫苗诱导小鼠细胞免疫应答的影响  

Effect of aluminum hydroxide adjuvant on cellular immune response induced with inactivated enterovirus 71 vaccine in mice

作　　者：王一平[1,2] 李玫颖[1,2] 高帆[1] 邵杰[1,3] 毛群颖[1] 姚昕[1] 程刚[2] 游松[2] 梁争论[1] 

机构地区：[1]中国食品药品检定研究院,北京100050 [2]沈阳药科大学,沈阳110016 [3]河南师范大学生命科学院,河南新乡453007

出　　处：《中国生物制品学杂志》2012年第8期939-942,共4页Chinese Journal of Biologicals

基　　金：国家科技支撑项目(2008BAI69B01);国家"十一五"传染病重大专项课题(2009ZX10004-804)

摘　　要：目的评价氢氧化铝佐剂对肠道病毒71型(Enterovirus 71,EV71)灭活疫苗诱导小鼠细胞免疫应答的影响。方法用含铝佐剂及无佐剂EV71灭活疫苗免疫BALB/c小鼠,并设生理盐水对照组。于加强免疫后7 d采血,进行小鼠脾单个核细胞(Mononuclear cell,MNC)的分离及CD8+T细胞的去除,采用酶联免疫斑点法(ELISPOT)检测经EV71疫苗原液或VP2-36刺激后的小鼠脾MNC分泌IFNγ水平;应用液相芯片技术(LUMINEX)检测经EV71疫苗原液刺激后的小鼠脾MNC分泌IL-2、IL-4、IL-5、IL-6和IL-10水平;采用体外微量中和试验法检测小鼠血清中和抗体效价。结果 BALB/c小鼠经2次免疫后,铝佐剂疫苗组小鼠脾MNC的IFNγSFC值显著高于无佐剂疫苗组(P均<0.05);铝佐剂疫苗组分泌IL-6和IL-10水平显著高于无佐剂疫苗组(P均<0.05);铝佐剂与无佐剂疫苗组小鼠血清EV71中和抗体阳转率均达到100%,铝佐剂疫苗组诱导的中和抗体效价显著高于无佐剂疫苗组(P<0.01)。结论 EV71灭活疫苗可诱导BALB/c小鼠产生特异性IFNγ、IL-2、IL-6和IL-10应答,铝佐剂可同时增强EV71灭活疫苗的Th1和Th2类免疫应答。Objective To evaluate the effect of aluminum hydroxide adjuvant on cellular immune response induced with inactivated enterovirus 71（EV71）in mice.Methods BALB / c mice were immunized with aluminum hydroxide adjuvant-containing and adjuvant-free inactivated EV71 vaccines respectively,using physiological saline as control.Blood samples were collected 7 d after booster immunization,from which mononuclear cells（MNCs） were isolated and CD8＋ T cells were removed.The secretion level of IFNγ by mouse splenic MNCs after stimulation with bulk EV71 vaccine or VPI-36 was determined by ELISPOT,while those of IL-2,IL-4,IL-5,IL-6 and IL-10 by LUMINEX technique.The serum neutralizing antibody titers of mice were determined by micro-neutralization test in vitro.Results The number of spot-forming cells（SFCs）of IFNγ secreted by MNCs of BALB / c mice immunized twice with aluminum hydroxide adjuvant-containing EV71 vaccine were significantly higher than those with adjuvant-free vaccine（P 〈 0.05）.Both the positive conversion rates of neutralizing antibody against EV71 in sera of mice immunized with aluminum hydroxide adjuvantcontaining and adjuvant-free vaccines were 100%.However,the neutralizing antibody titer induced by aluminum hydroxide adjuvantcontaining was significantly higher than that by adjuvant-free vaccine（P 〈 0.01）.Conclusion Inactivated EV71 vaccine induced secretion of specific IFNγ,IL-2,IL-6 and IL-10,and aluminum hydroxide adjuvant enhanced Th1 and Th2 immune responses induced by inactivated EV71 vaccine.

关 键 词：肠道病毒A型 人 氢氧化铝 佐剂 细胞免疫 

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