氢氧化锌胶体与三磷酸腺苷二钠联用对狂犬病疫苗诱导小鼠体液免疫应答的增强作用  

Enhancement of humoral immune response induced by rabies vaccine in mice by zinc hydroxide colloid combined with disodium adenosine triphosphate

作　　者：张浩[1] 乌美妮[1] 王海漩[1] 胡凝珠[1] 胡云章[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疫苗研究室云南省重大传染病重点实验室,昆明650118

出　　处：《中国生物制品学杂志》2012年第9期1115-1120,共6页Chinese Journal of Biologicals

基　　金：国家"艾滋病和病毒性肝炎等重大传染病防治"重大专项(2009ZX10004-702);国家高技术研究发展计划(863计划)(2012AA02A406);昆明市科技计划项目(昆科计字09H100302)

摘　　要：目的制备氢氧化锌[Zn(OH)2]胶体,并检测其单独或与三磷酸腺苷二钠(Disodium adenosine tripho-sphate,简称ATP)联用对狂犬病疫苗诱导小鼠体液免疫应答的增强作用。方法采用葡萄糖酸锌作为锌源,与NaOH反应制备Zn(OH)2胶体溶液,对其进行纯化及检测,并进行小鼠急性毒性试验;将不同剂量的Zn(OH)2胶体单独或与不同剂量的ATP联用,与狂犬病疫苗(0.25 IU)混合,免疫ICR小鼠,并设生理盐水对照组和抗原对照组,每组8只,Zn(OH)2和Zn(OH)2+ATP和生理盐水对照组采用单次免疫,抗原对照组分别免疫1、2、3、5次,分别于初次免疫后4、8、12、16周采血,分离血清,检测血清中抗狂犬病病毒特异性IgG抗体水平。结果所制备Zn(OH)2胶体溶液最大吸收波长为213 nm,可观察到明显的丁达尔现象,浓度为6 mg/ml,经光学色差显微镜及扫描电镜观察合格,小鼠急性毒性试验未观察到急性毒副作用。大部分佐剂组抗体水平高于抗原单次免疫组,但均未达到抗原5次免疫组水平;0.27和0.21 mg Zn(OH)2组在初次免疫后8周内有高水平抗体产生;0.27 mg Zn(OH)2组在初次免疫后16周出现抗体水平反弹性升高;0.18 mg Zn(OH)2+1.0 mg ATP组和0.18 mg Zn(OH)2+0.5 mg ATP组在初次免疫后4周内产生的抗体水平较高,之后逐渐下降;2次和3次免疫抗原对照组在初次免疫后8周内,其抗体水平不及佐剂组。结论所制备的Zn(OH)2胶体佐剂单独或与ATP联用均可增强狂犬病疫苗诱导小鼠特异性体液免疫应答,且安全性良好,有望应用于疫苗生产。Objective To prepare zinc hydroxide [Zn（OH）2] colloid and investigate the enhancement of humoral immune response induced by rabies vaccine in mice by the colloid alone or combined with disodium adenosine triphosphate（shortened as ATP）.Methods Zinc hydroxide colloid solution was prepared by the reaction of zinc gluconate and sodium hydroxide,then purified,identified and subjected to acute toxicity test in mice.ICR mice were divided into various groups,8 for each,and immunized with 0.25 IU rabies vaccine using the prepared colloid alone or combined with disodium ATP,at various dosages,as adjuvant.Physiological saline and antigen controls were set up.The mice in Zn（OH）2,Zn（OH）2 ＋ ATP and physiological saline control groups were immunized once,while those in antigen control groups were immunized for 1,2,3 and 5 times respectively.Blood samples were collected 4,8,12 and 16 weeks after the first immunization respectively,from which sera were separated and determined for specific IgG levels against rabies virus.Results The maximum absorbance wavelength of prepared zinc hydroxide colloid was 213 nm,at which obvious Tyndall phenomenon was observed.The colloid was qualified in optical and scanning electron microscopy,while showed no acute adverse reaction in mice.The antibody levels in most of Zn（OH）2 and Zn（OH）2 ＋ ATP groups were higher than those induced by a single dose of antigen.However,all the antibody levels induced by the vaccine containing adjuvant were lower than those by antigen for 5 times.High antibody levels were induced within 8 weeks after immunization with the vaccine containing 0.27 and 0.21 mg Zn（OH）2.The antibody level induced by the vaccine containing 0.27 mg Zn（OH）2 increased again 16 weeks after the first immunization.The antibody levels in 0.18 mg Zn（OH）2 ＋ 1.0 mg ATP and in 0.18 mg Zn（OH）2 ＋ 0.5 mg ATP groups were high within 4 weeks after the first immunization then decreased gradually.Within 8 weeks after the first immunization,the antibody levels i

关 键 词：氢氧化锌 胶体 佐剂 三磷酸腺苷二钠 狂犬病疫苗 体液免疫应答 

分 类 号：R392-33[医药卫生—免疫学]