低分子量肝素钠佐剂对甲型肝炎病毒疫苗诱导小鼠体液免疫应答的影响  

Effects of low molecular weight heparin sodium adjuvant on humoral immune response induced by Hepatitis A virus vaccine in mice

作　　者：王玮[1] 余柄廷 李建芳[1] 李彦涵[1] 刘丽君[3] 胡凝珠[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,昆明650118 [2]河北大学生命学院,河北保定071000 [3]昆明医科大学,昆明650500

出　　处：《中国比较医学杂志》2016年第1期19-24,共6页Chinese Journal of Comparative Medicine

基　　金：国家科技支撑计划项目(2014BAI01B01);云南省社会发展新药专项(2013BC011)

摘　　要：目的探讨低分子量肝素钠(low molecular weight heparin sodium,LMWH-Na)佐剂对甲型肝炎病毒(Hepatitis A virus,HAV)疫苗诱导小鼠体液免疫应答的影响。方法将ICR小鼠随机分成8组,6只/组,分别为甲型肝炎减毒活疫苗Hep A-l(18 EU)与5种不同剂量(100μL,20μL,10μL,1μL,0.1μL)的低分子量肝素钠混合免疫小鼠作为实验组,并取生理盐水作为阴性对照组(Blank)、Hep A-l(18 EU)作为阳性对照组、Hep A-l混合Al(OH)3(300μg)作为铝佐剂对照组,共免疫一次,于免疫后第4、8、12、16周进行尾静脉采血,采用ELISA法检测小鼠血清中抗HAV抗原特异性Ig G抗体水平。在试验期间,对小鼠的健康状况进行观察,并于免疫16周后,取低分子量肝素钠最大剂量组100μL组及生理盐水组小鼠肾脏、脾脏、肝脏、肺脏、心脏各主要器官制作病理切片进行对比观察。结果除阴性对照组外,各组小鼠免疫后第4周后均产生抗-HAV Ig G,并随时间的延长抗体水平逐渐升高,于第8周达到最高值,此后逐渐下降。同一时间不同组的抗体水平不同。其中,第4周,第8周,铝佐剂对照组,低分子量肝素钠100μL组,低分子量肝素钠20μL组,低分子量肝素钠10μL组的抗体水平高于阳性对照组且差异具有统计学意义(P<0.05),第12周,铝佐剂对照组,低分子量肝素钠20μL组和低分子量肝素钠10μL组的抗体水平高于阳性对照组(P<0.05),第16周,铝佐剂对照组和低分子量肝素钠10μL组的抗体水平高于阳性对照组(P<0.05),但在整个实验期间,低分子量肝素钠各组的抗体水平都低于铝佐剂对照组的抗体水平。表明低分子量肝素钠可增强HAV抗原特异性体液免疫应答,具有佐剂效应,最佳剂量为10μL/组,但其佐剂效应低于铝佐剂效应。试验期间,各组小鼠均未出现异常。低分子量肝素钠100μL组小鼠肾、脾、肝、肺、心组织均未观察到病变。结论低分子量肝素钠可明显增强HAV抗原诱导小鼠的�Objective To investigate the effect of Low molecular weight heparin sodium（ LMWH-Na） adjuvant onhumoral immune response induced by hepatitis A virus（ HAV） vaccine in mice. Methods ICR mice were randomly divided into 8 groups,six for each,and the mouse of each group are injected respectively with live attenuated hepatitis A vaccine Hep A-1（ 18 EU） ＋ different doses（ 100 μL,20 μL,10 μL,1 μL,0. 1 μL） of LMWH-Na,while those injected with physiological saline as a negative control group（ Blank）,those injected with Hep A- 1（ 18 EU） as positive control group,those injected with Al（ OH）3（ 300 μg） ＋ Hep A-1（ 18 EU） as the aluminium adjuvant control group,200μL for each. The specific Ig G level against HAV in sera of mice at weeks 4,8,12 and 16 after immunization were determined by indirect ELISA. During the experiment,the health status of the mice were observed,the kidney,spleen,liver,lung and heart of mice immunized with LMWH-Na 100 μL at the maximum dosage and those in negative control group were taken out at week 16 after immunization,and prepared into sections for pathological observation. Results In addition to the negative control group,mice in various group produced anti-HAV Ig G at 4 weeks after the immunization,of which the levels increased as time went on,and reached peak values at week 8 then decreased gradually. At 4 week and 8 week,the anti-HAV Ig G levels of aluminum adjuvant control group,LMWH-Na 100 μL group,LMWH-Na 20μL group,LMWH-Na 10μL group were higher than the positive control group and the difference was statistically significant（ P 〈0. 05）. At 12 week,the anti-HAV Ig G levels of aluminum adjuvant control group,LMWH-Na 20 μL group and LMWH-Na 10 μL group were higher than the positive control group and the difference was statistically significant（ P 〈0. 05）. 16 week,the anti-HAV Ig G levels of aluminum adjuvant control group and LMWH-Na 10 μL group were higher than positive control group and the difference was statistically significant

关 键 词：低分子量肝素钠 佐剂 甲型肝炎减毒活疫苗(HepA-l) 体液免疫 

分 类 号：R-332[医药卫生]