树鼩经甲型肝炎减毒活疫苗、乙型肝炎疫苗及联合硫酸乙酰肝素佐剂免疫后的免疫效果评价
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摘要
目的探讨甲型肝炎减毒活疫苗(HepA-1)、乙型肝炎疫苗(Hbv)以及HepA-1、Hbv分别与佐剂硫酸乙酰肝素(HS)联合对树鼩进行免疫后树鼩体液免疫应答的影响。方法按照疫苗种类、佐剂以及免疫途径将树鼩随机分组,同时设空白对照组,分别于末次免疫后的4周、8周、12周、16周和20周尾静脉采血,用ELISA法检测树鼩血清中的特异性IgG抗体水平。结果除空白对照组外,实验组均在末次免疫的第四周检测到抗甲型肝炎病毒(HAV)及抗乙型肝炎病毒(HBV)抗体,抗体水平随着免疫时间延长而逐渐升高并于12周达到峰值,此后逐渐下降。同时,联合HS佐剂组的体液免疫效果均优于单纯疫苗免疫组。并且不同抗原以及联合佐剂采用不同的免疫途径免疫树鼩其免疫效果存在显著性差异。结论通过对树鼩进行疫苗免疫原性和增强免疫效果检测,证实以树鼩作为动物模型进行疫苗评价的可行性。
Objective To investigate the humoral immune response of tree shrew after the immunization of Hepatitis A attenuated live vaccine(HepA-1)> Hepatitis B vaccine(Hbv) independently and combined with adjuvant heparan sulfate(HS).Methods The tree shrews were randomly divided into 10 groups by the type of vaccine,adjuvant and schedules.The specific IgG levels in sera of tree shrew were determined by ELISA at 4,8,12,16 and 20 weeks after the last immunization respectively.Results Except the blank control group,the anti-hepatitis A virus(HAV) and anti-hepatitis B virus(HBV) antibody was detected in all the experimental group at 4 weeks after last immunization,the level of immune antibody were gradually increased with time and reached the peak at the 12 weeks,then decreased gradually.At the same time,the humoral immunity effect of HS adjuvant group were better than that of vaccine group.There is a significant difference on immunity among tree shrews with different antigens and adjuvant by different immunization schedules.Conclusion The immune effect of vaccine immunogenicity and immune enhancing effect on tree shrew confirms the feasibility of the tree shrew as an animal model.
Objective To investigate the humoral immune response of tree shrew after the immunization of Hepatitis A attenuated live vaccine(HepA-1)> Hepatitis B vaccine(Hbv) independently and combined with adjuvant heparan sulfate(HS).Methods The tree shrews were randomly divided into 10 groups by the type of vaccine,adjuvant and schedules.The specific IgG levels in sera of tree shrew were determined by ELISA at 4,8,12,16 and 20 weeks after the last immunization respectively.Results Except the blank control group,the anti-hepatitis A virus(HAV) and anti-hepatitis B virus(HBV) antibody was detected in all the experimental group at 4 weeks after last immunization,the level of immune antibody were gradually increased with time and reached the peak at the 12 weeks,then decreased gradually.At the same time,the humoral immunity effect of HS adjuvant group were better than that of vaccine group.There is a significant difference on immunity among tree shrews with different antigens and adjuvant by different immunization schedules.Conclusion The immune effect of vaccine immunogenicity and immune enhancing effect on tree shrew confirms the feasibility of the tree shrew as an animal model.
引文
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[14]李海燕,黎家敏,李靖潇,等.高糖高脂饲料联合地塞米松诱发树鼩血糖、血脂异常[J].实验动物与比较医学,2010,30(3):197-200.
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[16]江勤芳,匡德宣,仝品芬,等.树鼩规模化繁殖及繁殖群建立[J].实验动物科学,2010,28(6):35-38.
[17]兰芸,王海漩,乌美妮,等.内源性危险信号低分子量硫酸乙酰肝素的免疫佐剂作用[J].中国生物制品学杂志,2011,24(1):5-9.
[18]施建东,胡凝珠,赵蕊蕊,等.氢氧化锌与低分子量透明质酸复合佐剂对甲肝乙肝联合抗原的体液免疫增强作用[J].中国生物制品学杂志,2012,25(1):34-36.
[19]梁亮,李嫒,岳慧芬,等.用人工繁殖的围生期和幼年期树鼩感染人乙型肝炎病毒的初步研究[J].中国人兽共患病学报,2006,22(8):792-795.
[20]Powell JD,Horton MR.Threat matrix:low-molecular-weight hyaluronan(HA)as a danger signal[J].Immunol Res,2005,31(3):207-218.
[2]Li S A,LEE WH,Zhang Y.Two bacterial infection models in tree shrew for evaluating the efficacy of antimicrobial agents[J].Zool Res,2012,33(l):l-6.
[3]Tong Y,Zhu Y,Xia Y,et al.Tupaia CD81,SR-BI,claudin,and occludin support hepatitis C virus infection[J].Virol J,.2011,85(6):2793-2802.
[4]黄晓燕,徐娟,孙晓梅,等.树鼩在人类疾病动物模型中应用研究进展[J],实验动物科学,201 3,30(2):59-64.
[5]Bukh J,Lanford RE,Purcell RH.Persistent human hepatitis B virus infection in cynomolgus monkeys:a novel animal model in the search for a cure[J].Hepatology,2013,58(5):1533-1536.
[6]Wang Q,Schwarzenberger P,Yang F,et al.Experimental chronic hepatitis B infection of neonatal tree shrews(Tupaia belangeri chinensis):a model to study molecular causes for susceptibility and disease progression to chronic hepatitis in humans[J].Virol J,2012,9:170.
[7]严瑞琪,苏建家,陈志英,等.人乙型肝炎病毒实验感染成年树鼩的初步研究[J].广西医学院学报,1984,1(1):10-15.
[8]Wang BJ,Tian YJ,Meng ZJ,et al.Establishing a new animal model for hepadnaviral infection:susceptibility of Chinese Marmota species to woodchuck hepatitis virus infection[J].Gen Virol,2011,92(3):681-691.
[9]He W,Ren B,Mao F,et al.Hepatitis D virus infection of mice expressing human sodium taurocholate co-transporting polypeptide[J].PLoS Pathog,2015,11(4):el004840.
[10]Bility MT,Cheng L,Zhang Z,et al.Hepatitis B virus infection and immunopathogenesis in a humanized mouse model:induction of human-specific liver fibrosis and M2-like macrophages[J].PLoS Pathog,2014,10(3):el004032.
[11]Walter E,Keist R,Niederost B,et al.Hepatitis infection of tupaia hepatocytes in vitro and in vivo[J].Hepatology,1996,24(l):l-5.
[12]Fan Y,Huang ZY,Cao CC,et al.Genome of the Chinese tree shrew[J].Nat Commun,2013,4:1426.
[13]Fan Y,Yu D,Yao YG.Tree shrew database(Tree shrew DB):agenomic knowledge base for the Chinese tree shrew[J].Sci Rep,2014,4:7145.
[14]李海燕,黎家敏,李靖潇,等.高糖高脂饲料联合地塞米松诱发树鼩血糖、血脂异常[J].实验动物与比较医学,2010,30(3):197-200.
[15]Meng BH,Liu H,Liang Y,et al.Investigation of pathogenesis of diabetic myopathy of skeletal muscle in experimental type I diabetes mellitus of tree shrews[J].Chin J Diabetes,2005,13(4):290-292.
[16]江勤芳,匡德宣,仝品芬,等.树鼩规模化繁殖及繁殖群建立[J].实验动物科学,2010,28(6):35-38.
[17]兰芸,王海漩,乌美妮,等.内源性危险信号低分子量硫酸乙酰肝素的免疫佐剂作用[J].中国生物制品学杂志,2011,24(1):5-9.
[18]施建东,胡凝珠,赵蕊蕊,等.氢氧化锌与低分子量透明质酸复合佐剂对甲肝乙肝联合抗原的体液免疫增强作用[J].中国生物制品学杂志,2012,25(1):34-36.
[19]梁亮,李嫒,岳慧芬,等.用人工繁殖的围生期和幼年期树鼩感染人乙型肝炎病毒的初步研究[J].中国人兽共患病学报,2006,22(8):792-795.
[20]Powell JD,Horton MR.Threat matrix:low-molecular-weight hyaluronan(HA)as a danger signal[J].Immunol Res,2005,31(3):207-218.