王陈芸;孙静;李萍;余柄廷;李建芳;李彦涵;王海漩;乌美妮;胡凝珠;胡云章;
WANG Chen-yun;SUN Jing;LI Ping;YU Bing-ting;LI Jian-fang;LI Yan-han;WANG Hai-xuan;WU Mei-ni;HU Ning-zhu;HU Yun-zhang;Institute of Medical Biology, Chinese Academy of Medical Science and Peking Union Medical College, Yunnan Key Laboratory of Vaccine Research & Development on Severe Infectious Diseases, Yunnan Research Center of Engineering Technique for Vaccines against Severe Infectious Diseases;

• 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疫苗研发重点实验室云南省重大传染病疫苗工程技术研究中心
• 2:河北大学生命学院

摘要(Abstract)：

目的探讨硫酸软骨素B(chondroitin sulfate B,CSB)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,以评价其佐剂效果。方法将ICR小鼠随机分成9组:甲型肝炎减毒活疫苗Hep A-l18 EU+不同剂量CSB(5、25、45、65、85、105μg)组、阴性对照组(生理盐水)、抗原对照组(Hep A-l 18 EU)和铝佐剂对照组[Hep A-l 18 EU+Al(OH)3300μg],每组8只,各组均经皮下注射ICR小鼠,200μl/只。于免疫后的第4、8、12和16周,采用间接ELISA法检测小鼠血清中抗-HAV特异性Ig G抗体水平。在实验期间,观察小鼠的健康状况,并于免疫16周后,取CSB 65μg组及阴性对照组小鼠肾脏、脾脏、肝脏、肺脏、心脏各主要器官,制备病理切片,进行对比观察。结果各组小鼠免疫后第4、8、12、16周,均产生抗-HAV Ig G抗体,除阴性对照组外,抗体水平均随时间的延长逐渐升高,于第8周达峰值,此后逐渐下降。CSB 65μg组抗体可长时间维持在高水平,至免疫后第12、16周,均高于抗原对照组(P<0.05),略高于铝佐剂对照组,最佳浓度为65μg/只。实验期间,各组小鼠均未出现异常;CSB65μg组小鼠肾脏、脾脏、肝脏、肺脏、心脏组织均未观察到病变。结论 CSB可明显增强HAV抗原诱导小鼠的体液免疫应答,具有成为新型疫苗佐剂的研发价值。
Objective To investigate the effect of chondroitin sulfate B(CSB) adjuvant on humoral immune response induced by hepatitis A virus(HAV)vaccine in mice. Methods ICR mice were randomly divided into nine groups, eight for each. The mice in six test groups were injected s. c. with live attenuated hepatitis A vaccine Hep A-l 18 EU + CSB at concentrations of 5, 25, 45, 65, 85 and 105 μg respectively, while those in negative control group with physiological saline, those in antigen control group with Hep A-l 18 EU alone, and those in aluminum control groups with Hep A-l 18 EU +300 μg aluminum hydroxide, 200 μl for each. The specific Ig G level against HAV in sera of mice at weeks 4, 8, 12 and16 after immunization were determined by indirect ELISA. The healthy statuses of mice were observed during the study.The kidney, spleen, liver, lung and heart of mice of mice immunized with Hep A-l + 65 μg CSB and those in negative control group were taken out at week 16 after immunization, and prepared into sections for pathological observation.Results Anti-HAV Ig Gs were induced in mice in various groups at weeks 4, 8, 12 and 16, of which the levels, except that in negative control group, increased as time went on, and reached peak values at week 8 then decreased gradually. The anti-HAV Ig G of mice immunized with Hep A-l + 65 μg CSB maintained at a high level for a long time, which was significantly higher than that in antigen control group(P < 0. 05) and slightly higher than that in aluminum adjuvant control group at weeks 12 and 16 after immunization. The optimal dosage of CSB was 65 μg for each mouse. No abnormality was observed in mice of various groups, while no pathological changes in the kidney, spleen, liver, lung and heart of mice immunized with Hep A-l + 65 μg CSB. Conclusion CSB enhanced the humoral immune responses induced by HAV antigen in mice significantly, which might be developed as a novel adjuvant of vaccines.

关键词(KeyWords)： 硫酸软骨素B;佐剂;甲型肝炎病毒疫苗;体液免疫
Chondroitin sulfate B(CSB);Adjuvant;Hepatitis A virus vaccine;Humoral immunity

Abstract:

Keywords:

基金项目(Foundation): 2013年云南省高技术产业发展项目计划:基因工程药物关键技术研究及新产品开发应用;云南省社会发展新药专项“治疗2型糖尿病Ⅰ类新药生物制品RBP4临床前研究”(2013BC011)

作者(Authors): 王陈芸;孙静;李萍;余柄廷;李建芳;李彦涵;王海漩;乌美妮;胡凝珠;胡云章;
WANG Chen-yun;SUN Jing;LI Ping;YU Bing-ting;LI Jian-fang;LI Yan-han;WANG Hai-xuan;WU Mei-ni;HU Ning-zhu;HU Yun-zhang;Institute of Medical Biology, Chinese Academy of Medical Science and Peking Union Medical College, Yunnan Key Laboratory of Vaccine Research & Development on Severe Infectious Diseases, Yunnan Research Center of Engineering Technique for Vaccines against Severe Infectious Diseases;

DOI: 10.13200/j.cnki.cjb.000843

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