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目的评价融合基因pcDNA3.1/HisB-IL-2-G1-G2的DNA免疫效果。方法肌注免疫小鼠,分次采血,用酶联免疫法(ELISA)直接免疫酶斑减少中和试验检测小鼠体液免疫;淋巴细胞增殖试验(MTT法)检测其细胞免疫应答。免疫后第六周感染病毒。间接免疫荧光试验(IFIA)观察免疫小鼠抗病毒感染能力。结果pcDNA3.1/HisB-IL-2-G1-G2可诱导小鼠产生特异性抗汉坦病毒抗体和中和抗体,其效价均高于其他对照组,差异显著(P<0.05)。MTT实验表明,pcDNA3.1/HisB-IL-2-G1-G2诱导小鼠脾细胞对病毒抗原的增殖指数明显高于其他对照组。IFIA结果显示免疫小鼠对病毒攻击有保护性。结论pcDNA3.1/HisB-IL-2-G1-G2可诱导小鼠产生特异性体液和细胞免疫应答,为新一代汉坦病毒(HTV)DNA疫苗开发提供了实验依据。
Objective To evaluate the DNA immunization effect of fusion gene pcDNA3.1 / HisB-IL-2-G1-G2. Methods Mice were intramuscularly immunized and the blood was collected at different times. The humoral immunity of mice was detected by enzyme-linked immunosorbent assay (ELISA), and the cellular immune response was detected by MTT assay. Six weeks after immunization, the virus is infected. Indirect immunofluorescence assay (IFIA) was used to observe the ability of anti-virus in immunized mice. Results The pcDNA3.1 / HisB-IL-2-G1-G2 induced mouse anti-Hantavirus antibody and neutralizing antibody. The titer was higher than that of other control groups (P <0.05). MTT assay showed that the proliferation index of spleen cells induced by pcDNA3.1 / HisB-IL-2-G1-G2 was significantly higher than that of other control groups. IFIA results show that immunized mice are protective against viral challenge. Conclusion The pcDNA3.1 / HisB-IL-2-G1-G2 can induce specific humoral and cellular immune responses in mice, providing an experimental basis for the development of a new generation of Hantavirus (HTV) DNA vaccine.