硒化卡拉胶对甲型肝炎减毒活疫苗诱导的小鼠体液免疫应答的影响

doi:10.3969/j.issn.1674-5817.2018.02.006

摘要/Abstract

摘要： 目的研究硒化卡拉胶(KSC)对甲型肝炎减毒活疫苗(HepA-1)诱导的小鼠体液免疫应答的影响。方法随机分成7个组,每组7只ICR小鼠。分组: 阴性对照组(生理盐水)、单纯抗原组(18EU HepA-1)、铝佐剂组[Al(OH)₃ 0.3 mg+18EU HepA-1]、KSC实验组(KSC 1 mg+18EU HepA-1、KSC 0.5 mg+18EU HepA-1、KSC 0.25 mg+18EU HepA-1、KSC 0.1 mg+18EU HepA-1),皮下多点免疫注射,注射总体积300 µL,免疫1次。免疫注射后及其后4周、8周、12周、16周尾静脉采血,取血清用间接酶联免疫吸附法测定抗-甲型肝炎病毒(HAV) IgG抗体滴度。在实验期间,观察小鼠的健康状况,并于免疫后 16 周,取KSC最佳剂量组及阴性对照组小鼠肾脏、脾脏、肝脏、肺脏、心脏组织,制备病理切片,进行对比观察。结果除阴性对照组外,其余各组在各个检测时间点均检测到抗HAV IgG抗体, 且存在先升高后降低的趋势,铝佐剂组在12周达到峰值, 其余各组在8周达到峰值。KSC 0.5 mg组在8周、12周抗体水平高于单纯抗原组(t=3.103, t=2.573, P<0.05), KSC 0.25 mg组在12周抗体水平高于单纯抗原组(t=2.602,P<0.05)。本次实验的最佳剂量为KSC 0.5 mg。试验期内,观察小鼠各项生理指标未见异常,KSC 0.5 mg 组小鼠心脏、肝脏、脾脏、肺脏、肾脏组织均未观察到病变。结论 KSC能够作为佐剂增强HepA-1诱导的小鼠体液免疫反应。

关键词: 硒化卡拉胶(KSC), 佐剂, 甲型肝炎减毒活疫苗(HepA-l), 体液免疫

Abstract: Objective To investigate the effect of kappa-selenocarrageenan (KSC) on humoral immune response induced by hepatitis A virus attenuated live vaccine (HepA-1) in mice.Methods Seven experiment groups were set up,with 7 ICR mouse per group randomly,negative control group (saline),pure antigen group (18EU HepA-1),Aluminium adjuvant group [Al(OH)₃ 0.3 mg+18 EU HepA-1],KSC experimental groups (KSC 1 mg+18 EU HepA-1,KSC 0.5 mg+18 EU HepA-1,and KSC 0.25 mg +18 EU HepA-1,KSC 0.1 mg+18 EU HepA-1),in which mice were injected subcutaneously with 300 µL,immunizing one time.The anti-HAV specific IgG antibody levels were tested by indirect ELISA method in the serum of mice after the first 4th,8th,12th,16th week of injection.The healthy status of mice were observed during the study.The kidney,spleen,liver,lung and heart of mice immunized with KSC+HepA-l at the optimal dosage and those in negative control group were taken out at 16th week after immunization,and prepared into sections for pathological observation.Results HAV-specific IgG antibody of all mice were detected at four setting time point except negative control group.The antibody level of all groups increased first and then decreased.The peak of Aluminium adjuvant group at the 12th week and others at the 8th week.The level of antibody in KSC 0.5 mg+18EU HepA-1 group at 8th,12th week was higher than that in simple antigen group (t=3.103,t=2.573,P<0.05).The level of antibody in KSC 0.25 mg +18EU HepA-1 group at week 12 was higher than that in pure antigen group(t=2.602,P<0.05).The optimal dosage of mice in this test was 0.5 mg.No abnormality was observed in the physiological indexes of mice in various groups,while there was no pathological changes in the kidney,spleen,liver,lung and heart of mice immunized with HepA-l +0.5 mg KSC.Conclusion KSC can be used as an adjuvant to enhance humoral immune response induced by HepA-1 in mice.

Key words: Kappa-selenocarrageenan(KSC), Adjuvant, Hepatitis A virus attenuated live vaccine (HepA-1), Humoral immune

中图分类号:

Q95-33

贾森泉, 金晓, 李晓红, 王海璇, 胡凝珠, 胡云章. 硒化卡拉胶对甲型肝炎减毒活疫苗诱导的小鼠体液免疫应答的影响[J]. 实验动物与比较医学, 2018, 38(2): 111-116.

JIA Sen-quan, JIN Xiao, LI Xiao-hong, WANG Hai-xuan, HU Ning-zhu, HU Yun-zhang. Effect of Kappa-selenocarrageenan on Humoral Immune Response Induced by Hepatitis a Virus Attenuated Live Vaccine in Mice[J]. Laboratory Animal and Comparative Medicine, 2018, 38(2): 111-116.

参考文献

[1] Gherardi RK,Aouizerate J,Cadusseau J,et al.Aluminum adjuvants of vaccines injected into the muscle:Normal fate,pathology and associated disease[J].Morphologie,2016,100(329):85-94.
[2] Di R M.Biological properties of carrageenan[J].J Pharm Pharmacol,1972,24(2):89-102.
[3] Shu Y,Liu XB,Ma XH,et al.Immune response mechanism of mouse monocytes/macrophages treated with κ-carrageenan polysaccharide[J].Environ Toxicol Pharmacol,2017,53:191-198.
[4] Chan WI,Zhang G,Li X,et al.Carrageenan activates monocytes via type-specific binding with interleukin-8:an implication for design of immuno-active biomaterials[J].Biomater Sci,2017,5(3):403-407.
[5] Bhattacharyya S,Liu H,Zhang Z,et al.Carrageenan-induced innate immune response is modified by enzymes that hydrolyze distinct galactosidic bonds[J].J Nutr Biochem,2010,21(10):906-913.
[6] 周革非,李树福,王长海.κ-卡拉胶硫酸多糖的免疫调节活性初步研究[J].海洋科学,2010,34(8):56-59.
[7] 陶慧娜,孙涛,周冬香,等.卡拉胶及其衍生物的生物活性研究进展[J].安徽农业科学,2008,36(19):7967-7968.
[8] 卢岚,鲁兵,王春娥.保健食品中有机硒和无机硒的分析探讨[J].中国卫生检验杂志,2010,20(04):767-768.
[9] 崔乔,尚德静,邹霞.硒多糖的研究进展[J].中国生化药物杂志,2003,24(3):155-157.
[10] 张罗修,蒋建明,贾永锋.硒化卡拉胶对肿瘤坏死因子生成的影响(英文)[J].中国药学(英文版),1994,3(1):37-42.
[11] 俞银姣,张罗修.硒化卡拉胶对小鼠自然杀伤细胞活性的影响[J].中国临床药学杂志,1996,5(2):60-63.
[12] Ji YB,Ling N,Zhou XJ,et al.Schedule-dependent effects of kappa-selenocarrageenan in combination with epirubicin on hepatocellular carcinoma[J].Asian Pac J Cancer Prev,2014,15(8):3651.
[13] Mohan T,Verma P,Rao N.Novel adjuvants &delivery vehicles for vaccines development:a road ahead[J].Indian J Med Res,2013,138(5):779-795.
[14] Esch HH.Mechanism of immunopotentiation and safety of aluminum adjuvants[J].Front Immunol,2013,3(1):1-13.
[15] 孙丽莎,冯婷,路静,等.牛膝多糖与锌对小鼠树突状细胞增殖分化和抗原提呈能力的影响[J].中草药,2010,41(8):1319-1322.
[16] 韩洁,岳文鹏,何光志,等.八种中药多糖对小鼠巨噬细胞释放细胞因子的影响[J].现代免疫学,2011,31(6):495-498.
[17] 李婧文,周长林.多糖的免疫调节作用及多糖药物的研究进展[J].中国生化药物杂志,2016,36(4):24-28.
[18] 刘轶博,耿兴超,汪巨峰,等.免疫佐剂作用机制研究新进展[J].中国新药杂志,2015,24(20):2324-2329.
[19] 雍妍,王茹静,黄青,等.天然药物化学成分结构修饰研究进展[J].中药与临床,2015,6(06):55-60.
[20] 刘旭.板蓝根多糖及其硫酸化衍生物体外抗病毒和增强免疫活性的比较[D].南京:南京农业大学,2012.
[21] 吴欣,张纯东,戴敏.硒化卡拉胶口服液对免疫抑制小鼠免疫功能的影响[J].中国临床药理学与治疗学,2007,12(11):1269-1273.
[22] 王玮,余柄廷,李建芳,等.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[J].中国比较医学杂志,2016,26(1):19-24.