目的:构建结核杆菌ESAT6抗原DNA疫苗(pE)和泛素基因与ESAT6抗原基因融合的DNA疫苗(pUE)。方法:分别将构建的DNA疫苗肌内注射免疫BALB/c雌性小鼠,检测小鼠的血清抗体(IgGI、gG1、IgG2a)、细胞因子(IFN-γI、L-4)和细胞毒性T淋巴细胞反应(CTL),比较融合基因DNA疫苗和单基因DNA疫苗诱生免疫应答的强度。结果:pE组小鼠血清IgG水平高于pUE组(P〈0.01),但IgG2a/IgG1比值低于pUE组[(2.28±0.40)vs(3.87±0.60),P〈0.05]。与pE组相比,pUE组小鼠IFN-γ分泌水平增高(P〈0.01),IL-4分泌水平下降(P〈0.01);pUE增强了CTL活性。提示融合基因DNA疫苗诱生的抗原特异的体液免疫应答不及单基因DNA疫苗,但其能诱导更强的细胞免疫应答。结论:泛素-ESAT6融合基因DNA疫苗对于防治结核病可能比单基因DNA疫苗更为有效。
Objective:To construct Mycobacterium tuberculosis ESAT6 (pE) antigen DNA vaccine and ubiquitin-ESAT6 fusion gene (pUE) DNA vaccine. Methyls: The constructed DNA vaccines were intramuscularly inoculated into female BALB/c mice separately. The serum antibodies (including IgG, IgG1, and IgG2a ), Cytokines (IFN-γ and IL-4), and cytotoxic T lymphocytes response were detected in the immunized mice. Results: Mice in pE group had a higher serum level of IgG (P〈 0.01) and a lower value of IgG2a/IgG1 ([2.28±0.40] vs [3.87±0.60] ,P〈0.05) than mice in pUE group. Besides, mice in pUE group secreted more IFN-γ than those in pE group (P〈0.01), but secreted less IL-4 (P〈0.01). Furthermore, pUE enhanced the activity of CTL. The results showed that pUE DNA vaccine induced weaker humoral immune response, but stronger cellular immune responses compared to pE DNA vaccine. Conclusion.. The pUE DNA vaccine constructed in this study sheds new lights on the prophylactic and therapy of tuberculosis.