含PreS1、PreS2和S抗原的重组乙型肝炎疫苗辅以不同佐剂在HBV转基因小鼠中的免疫原性  

Immunogenicity of recombinant hepatitis B vaccine containing PreS1, PreS2 and S antigens with different adjuvants in HBV transgenic mice

作　　者：叶琳[1] 胡波[1] 张鹏艳 蒋丽明[1] 施慧敏[1] 谭昌耀 

机构地区：[1]成都生物制品研究所有限责任公司生物技术研究室,610023

出　　处：《国际生物制品学杂志》2017年第3期109-115,共7页International Journal of Biologicals

摘　　要：目的 研究以免疫刺激复合物（immune stimulating complex，ISCOM）或Al（OH）3为佐剂的含PreS1、PreS2和S抗原的重组乙型肝炎疫苗在HBV转基因小鼠中诱导的免疫应答。方法 将纯化的含PreS1、PreS2和S抗原的重组HBsAg（SS1S2）分别辅以ISCOM或Al（OH）3佐剂制成疫苗。HBV转基因小鼠每月肌内注射1针疫苗，共注射7针。免疫前及每针后1个月称量小鼠体重，观察小鼠生长情况。免疫前及每针后1个月采血，用ELISA检测HBV S、PreS1、PreS2抗原及其相应抗体。7针后1个月分离小鼠脾淋巴细胞，用酶联免疫斑点试验分别测定分泌IL-4、IL-5、IFN-γ的效应T细胞频数。结果 小鼠生长状况良好，每针后体重均有增长。首针后1个月，10只ISCOM＋SS1S2免疫鼠分别有3和1只抗S和PreS1抗体阳转，无一小鼠抗PreS2抗体阳转；10只Al（OH）3＋SS1S2免疫鼠的抗体均为阴性。7针后1个月，10只ISCOM＋SS1S2免疫鼠的抗S和PreS1抗体均阳转，8只抗PreS2 抗体阳转；10只Al（OH）3＋SS1S2免疫鼠分别有9、7和1只抗S、PreS1和PreS2 抗体阳转；ISCOM＋SS1S2组和Al（OH）3＋SS1S2组的抗S抗体几何平均滴度分别为1：15 647和1：1 039，差异有统计学意义（t=5.18，P=0.000）。7针后1个月，ISCOM＋SS1S2组和Al（OH）3＋SS1S2组分泌IL-4的斑点细胞数分别为219和78斑点形成细胞（spot-forming cell，SFC）/106细胞（t=10.50，P=0.000），分泌IL-5的斑点细胞数分别为286和34 SFC/106细胞（t=19.53，P=0.000 ），差异有统计学意义。结论 含PreS1、PreS2和S抗原的重组乙型肝炎疫苗辅以ISCOM或Al（OH）3佐剂在HBV转基因小鼠中均具有免疫原性，且ISCOM＋SS1S2的免疫原性强于Al（OH）3＋SS1S2。Objective To evaluate immune responses induced by recombinant hepatitis B vaccine containing PreS1, PreS2 and S antigens （SS1S2） with immune stimulating complex （ISCOM） or Al（OH）3 adjuvants in HBV transgenic mice. Methods ISCOM＋SS1S2 and Al（OH）3＋SS1S2 vaccines were made by combining purified SS1S2 antigen with ISCOM matrix or Al（OH）3 adjuvant. HBV transgenic mice were injected intramuscularly once a month for a total of 7 doses. The weight of each mouse was measured before immunization and 1 month after each injection, and the growth status of mice were observed. Blood was collected at same time. PreS1, PreS2 and S antigens and their corresponding mouse antibodies were detected by ELISA. The spleen cells of mice were separated and frequencies of IL-4, IL-5, IFN-γ-secreting cells were detected by enzyme-linked immunospot assay. Results Mice growth status was good, and body weight kept increasing after each injection. After the first injection, 3, 1 and 0 of 10 mice immunized with ISCOM＋SS1S2 had anti-S, anti-PreS1 and anti-PreS2 seroconversion, respectively; 10 mice immunized with Al（OH）3＋SS1S2 were all negative for antibodies . After 7 doses injection, 10, 10 and 8 of 10 mice immunized with ISCOM＋SS1S2 had anti-S, anti-PreS1 and anti-PreS2 seroconversion, respectively; 9, 7 and 1 of 10 mice immunized with Al（OH）3＋SS1S2 were anti-S, anti-PreS1 and anti-PreS2 seroconversion, respectively. The geometric mean titers of anti-S of ISCOM＋SS1S2 group and Al（OH）3＋SS1S2 group were 1：15 647 and 1：1 039 , respectively , and the difference between two groups was statistically significant . （t=5.18, P=0.000） After 7 doses injection, the frequencies of IL-4-secreting cell of ISCOM＋SS1S2 group and Al（OH）3＋SS1S2 group were 219 and 78 spot-forming cell （SFC）/106 cells, respectively （t=10.50, P=0.000）, and the frequencies of IL-5-secreting cell of ISCOM＋SS1S2 group and Al（OH）3＋SS1S2 group were 286 and 34 SFC/106 cells, respectively （t=19.53, P=0.

关 键 词：肝炎表面抗原 乙型 小鼠 转基因 免疫刺激复合物 免疫原性 

分 类 号：R392[医药卫生—免疫学]