阳离子脂质体DOTAP作为乙肝疫苗佐剂的免疫增强效果  

Immunopotentiation of cationic liposome DOTAP as an adjuvant of hepatitis B vaccine

作　　者：喻刚[1] 郝鹏亮[1] 韩锡鑫[1] 韩静[1] 赵巍[1] 黄晓媛[1] 杨晓明 

机构地区：[1]武汉生物制品研究所有限责任公司,湖北武汉430207 [2]国药集团中国生物技术股份有限公司,北京100029

出　　处：《中国生物制品学杂志》2016年第10期1017-1020,1026,共5页Chinese Journal of Biologicals

基　　金：2013年国家新药创制项目"疫苗综合性技术研究开发大平台建设"子课题:PDS纳米佐剂疫苗(2013ZX09402302-229);2014年国家新药创制项目"新型纳微颗粒佐剂及黏膜免疫佐剂的研究"子课题:脂质体纳米粒的研究(2014ZX09102045-011)

摘　　要：目的探讨阳离子脂质体DOTAP（2,3-二油氧基丙基三甲基氯化铵）作为乙肝疫苗佐剂的免疫增强效果。方法薄膜蒸发复水法制备DOTAP阳离子脂质体乙肝疫苗,检测其粒径、Zata电位、包封率;将BALB/c小鼠随机分为5组：DOTAP阳离子脂质体乙肝疫苗组（DOTAP佐剂组）、不加佐剂的乙肝疫苗组（HBS组）、乙肝疫苗加铝佐剂组（铝佐剂组）、乙肝疫苗加CPG-ODN佐剂组（CPG-ODN佐剂组）以及PBS对照组,免疫剂量分别为0.2、2μg/（只·0.1ml）,每个剂量10只小鼠,分别于0、28d经肌肉免疫,0、14、28、42d采血,分离血清,ELISA法检测血清中抗HBs特异性IgG抗体、亚型抗体（IgG1、IgG2a）滴度。结果制备的阳离子脂质体乙肝疫苗平均粒径200~300nm,粒径分布均匀,带正电荷,包封率在70%以上。DOTAP佐剂组血清IgG抗体水平明显高于HBS组,其中2μg剂量组差异有统计学意义（P〈0.05）;与铝佐剂组相比,两个剂量组差异均无统计学意义（P〉0.05）;0.2μg剂量组显著低于CPG-ODN佐剂组（P〈0.05）。DOTAP佐剂组可同时诱导高水平的IgG1、IgG2a型抗体,两个剂量组产生的IgG1型抗体水平均与铝佐剂组相当（P〉0.05）,但IgG2a型抗体滴度均明显高于铝佐剂组（P〈0.05）;两个剂量DOTAP佐剂组产生的IgG1、IgG2a型抗体水平与CpG-ODN佐剂组相比,差异均无统计学意义（P〉0.05）。结论DOTAP阳离子脂质体作为重组乙肝疫苗的佐剂具有良好的免疫增强效果,其对体液免疫的增强作用不劣于铝佐剂和CPG-ODN佐剂,DOTAP佐剂可同时诱导Th2细胞和Th1细胞介导的免疫应答。Objective To investigate the immunopotentiation of cationic liposome DOTAP（2, 3- dioleoyloxy- propyl-trimethylammonium） as an adjuvant of hepatitis B（HB） vaccine. Methods DOTAP cationic liposome HB vaccine was prepared by lipid film evaporating and hydrating, of which the size, Zata potential and encapsulation efficiencies were measured. BALB/c mice were randomly divided into DOTAP adjuvant, HBS, aluminum adjuvant, CPG-ODN adjuvant and PBS control groups, and inoculated i. m. with HB vaccine containing DOTAP cationic liposome adjuvant, adjuvant-free HB vaccine, HB vaccine containing aluminum adjuvant, HB vaccine containing CPG-ODN adjuvant and PBS respectively.The dosages of vaccines were 0. 2 and 2 μg / 0. 1 ml respectively, and each dosage was inoculated to 10 mice for 2 times on days 0 and 28. Serum samples were collected on days 0, 14, 28 and 42, and determined for Ig G, IgG1 and IgG2a titers against HBs. Results The sizes of prepared cationic liposome HB vaccine was 200 ~ 300 nm, which were distributed evenly. The encapsulation efficiency of the vaccine, with positive charges, reached more than 70%. The anti-HBs Ig G titers in of DOTAP adjuvant group was higher than that in HBs group, of which that induced by 2 μg of vaccines showed significant difference（P〈0. 05）. However, the Ig G titers induced by 0. 2 and 2 μg of HB vaccine containing DOTAP adjuvant showed no significant difference with those by the vaccine containing aluminum adjuvant at the corresponding dosages（P〉0. 05）, while that induced by 0. 2 μg of vaccine containing DOTAP adjuvant was significantly lower than that by the vaccine containing Cp G-ODN adjuvant at the same dosage（P〈0. 05）. High IgG1 and IgG2a titers were induced in DOTAP adjuvant group. The IgG1 titers induced by the vaccine containing DOTAP adjuvant at two dosages were equivalent to（P〉0. 05）, while the IgG2a titers were significantly higher than（P〈0. 05）, those by the vaccine containing aluminum adjuvant. However, both IgG1 and IgG2a titers

关 键 词：2 3-二油氧基丙基三甲基氯化铵 阳离子脂质体 乙肝疫苗 佐剂 抗体 

分 类 号：R373.21[医药卫生—病原生物学] R392-33[医药卫生—基础医学]