乙肝疫苗联合含CpG基序寡脱氧核苷酸和卡介苗对小鼠的免疫应答影响

Effect of hepatitis B vaccine with CpG oligodeoxynucleotide or Bacilli Calmette Guerin on immune responses in mice

目的:观察乙肝疫苗联合CpGODN和卡介苗免疫小鼠,对小鼠抗HBs水平和细胞免疫的影响.方法:不同剂量乙肝疫苗单用或联合CpGODN或卡介苗免疫小鼠,用ELISA方法检测抗HBs水平,流式细胞仪检测外周血CD4+和CD8+T淋巴细胞亚群的百分比和比值.结果:免疫4wk后,中、高剂量乙肝疫苗联合CpGODN组抗HBs水平显著高于低剂量乙肝疫苗组(420.56±55.66vs181.62±41.25,t=0.013,P<0.05;403.38±63.85vs181.62±41.25;t=0.024,P<0.05).但中、高组间无差别.CpGODN联合组与单用乙肝疫苗组比较,联合低、中剂量乙肝疫苗抗HBs水平较单用乙肝疫苗明显升高(181.62±41.25vs59.16±18.43,t=0.022,P<0.05;420.56±55.66vs241.82±23.84,t=0.018,P<0.05).但高剂量乙肝疫苗联合组与单用乙肝疫苗比较两组间未显示明显差别.且以CpGODN联合中等量乙肝疫苗产生的抗HBs水平最高.卡介苗联合组与单用乙肝疫苗组无差别,但在联合低、中剂量乙肝疫苗抗HBs水平低于CpGODN联合组.CpGODN联合组CD4+T淋巴细胞百分比和CD4+/CD8+比值明显高于单用乙肝疫苗组和卡介苗联合组(CD4+:38.30±1.68vs31.47±2.15,t=0.018,P<0.05;38.30±1.68vs29.31±2.97,t=0.013,P<0.05;CD4+/CD8+:9.01±0.38vs6.45±0.39,t=0.000,P<0.05;9.01±0.38vs6.99±0.79,t=0.029,P<0.05).结论:CpGODN联合乙肝疫苗不仅能诱导出高水平的抗HBs,而且对细胞免疫也有调节作用.

AIM： To investigate the effects of hepatitis B vaccine with CpG oligodeoxynucleotide （ODN） or Bacilli Calmette Guerin on the immune responses in mice. METHODS： BALB/c mice were subcutaneously injected with different doses of hepatitis B vaccine （0.25, 0.75, 1.0μg） combined with CpG ODN or BCG. The levels of serum hepatitis surface antibody （anti-HBs） were detected by ELISA assays two and four weeks after injection. The changes of peripheral blood T lymphocyte subsets CD4^±, CD8^± and CD4^±/CD8^± ratio were detected by flow cytometry. RESULTS： Four weeks after injection, the levels of serum anti-HBs in the mice injected with the combination of CpG ODN and hepatitis B vaccine （0.75 and 1.0 μg, respectively） were significantly higher than that with CpG ODN and 0.25 μg hepatitis B vaccine （420.56±55.66 vs 181.62±41.25, t = 0.013, P 〈0.05; 403.38±63.85 vs 181.62±41.25; t = 0.024, P 〈0.05）. Compared with that in the mice only injected with hepatitis B vaccine, the serum anti-HBs levels in the ones injected with the combination of CpG ODN and hepatitis B vaccine （0.25 and 0.75μg, respectively） were markedly higher （181.62±41.25 vs 59.16±18.43, t = 0.022, P 〈0.05; 420.56±55.66 vs 241.82±23.84, t = 0.018, P 〈0.05）. The serum anti-HBs levels in the mice injected with the combination of BCG and hepatitis B vaccine were not significantly different from that with hepatitis B vaccine only. Whereas compared with that in the CpG ODN combination groups, the serum anti-HBs levels were lower in the mice injected with the combination of BCG and 0.25 or 0.75 μg hepatitis B vaccine. The percentages of CD4^＋ and the CD4^＋/CD8^＋ ratio in the mice immunized by CpG ODN combined with hepatitis B vaccine increased significantly as compared with those with hepatitis B vaccine only and the BCG combination group （CD4^＋ 38.30±1.68% vs 31.47±2.15%, t = 0.018, P 〈0.05; 38.30±1.68 vs 29.31±2.97, t = 0.013, P 〈0.05; CD4^＋/CD8^＋ 9.01 ±0.38 vs 6.45±0.39, t = 0.000, P 〈0.05; 9.01±0.38 vs 6.99±0.79, t = 0.029, P 〈0.05）. CONCLUSION： CpG ODN combined with hepatitis B vaccine not only induces high level of anti-HBs, but also enhances the cellular immune responses.