中国汉坦病毒H8205株G1-G2-人源IL-2融合基因DNA免疫的实验研究

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.

The Immune Responses of DNA Vaccination of Hantavirus H8205 Strain Glycoproteins G1, G2 and Human IL- 2 Fusion Gene

Objective To evaluate the immune responses of DNA vaccination of pcDNA3.1/HisB-IL-2-G1-G2fusion gene.Methods The Balb /c mice were genetically immunized intramuscularly(i.m).Blood samples were collected every two weeks.The humoral immune r esponse of the mice was determined by ELISA and plaque reduction neutrali zation test(PRNT).The cell immune response of the mice was determined by MTT method.The pro tective ability was measured by IFIA.Results pcDNA3.1/HisB-IL-2-G1-G2could elicit significant antibody response s and also producing neutralizing antibodies.The antibodies titer in duced by pcDNA3.1/HisB-IL-2-G1-G2is much higher than other groups(P<0.05).The result of MTT assay showed that th e cell immune response of pcDNA3.1/H isB-IL-2-G1-G2immunized mice was significantly higher than other gro ups.Conclusion DNA vaccination of pcDNA3.1/HisB-IL-2-G1-G2could induce thehumoral and cell immune responses.T his study provides a basis for the development of a new generation Hantavirus vaccine.