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

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+ SS1 S2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0=5.18,P=0.000).7针后1个月,ISCOM+ SS1S2组和Al(OH)3+ SS1S2组分泌IL-4的细胞频数分别为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的细胞频数分别为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+ SS1 S2的免疫原性强于Al(OH)3+ SS1S2.

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

Objective To evaluate imm une responses induced by recombinant hepatitis B vaccine containing PreS1,PreS2 and S antigens (SS1S2) with immune stimulating complex (ISCOM) or Al(OH)3adjuvants 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 was 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 had anti-S,anti-PreS1 and anti-PreS2 seroconversion,respectively.The geometric mean titers of anti-S in 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 =0.000).The difference between two groups was statistically significant.Conclusion ISCOM + SS1S2 and Al (OH)3 + SS1S2 are both immunogenic in HBV transgenic mice,and the immunogenicity of ISCOM + SS1S2 is higher than that of Al(OH)3 + SS1S2.