PTD-HBcAg融合蛋白诱导HBV特异性免疫反应的研究

The Experimental Study of Specific Immune Response Induced by PTD-HBcAg Fusion Protein

目的观察PTD-HBcAg融合蛋白免疫BALB/c小鼠后的体液免疫与细胞免疫效果。方法目的蛋白(PTD-HBcAg)、阳性对照蛋白(HBcAg)和阴性对照分别与等体积的弗氏完全佐剂乳化免疫小鼠,第21天以不完全弗氏佐剂乳化同法加强免疫,第28天ELISA法检测HBcAb滴度,流式细胞仪检测HBV特异性IFN-γ+、CD8+T淋巴细胞水平。结果PTD-HBcAg融合蛋白能够诱导产生较高水平HBcAb;与阳性蛋白对照组[(1.45±0.19)%,(2.19±0.04)%和(4.32±1.04)%]和阴性对照组相比,PTD-HBcAg融合蛋白组[(2.64±0.21)%,(3.50±0.50)%和(6.85±1.3)%]诱导的HBV特异性CD8+T淋巴细胞水平明显增高(P<0.05)。结论PTD-HBcAg融合蛋白具有体液免疫与细胞免疫效果,为治疗性乙肝疫苗的开发提供了一定的实验基础。

Objective To observe the humoral and cellular immune effects of PTD-HBcAg fusion protein in mice.Methods BALB/c mice were subcutaneously immunized at the tail base with 100 μl of freshly prepared emulsion containing PTD-HBcAg,HBcAg,PTD,PBS and an equal volume of CFA.Mice were equally booster immunized with the antigen mixture emulsified in IFA,21 days later.Sera were collected to test HBcAb and splenocytes were prepared to study IFN-γ＋,CD8＋ T lymphocytes on day 28.Results PTD-HBcAg fusion protein could induce higher levels of HBcAb;The amount of specific CD8＋ T lymphocytes induced by PTD-HBcAg was much more than that induced by others[（2.64±0.21）%,（3.50±0.50）%,（6.85±1.30）% vs（1.45±0.19）%,（2.19±0.04）%,（4.32±1.04）%;P0.05].Conclusion PTD-HBcAg fusion protein showed good humoral and cellular immune effects,which provided a experimental basis for the development of therapeutic hepatitis B vaccine.