【摘要】 目的探讨汉坦病毒H8205株G1人源IL2融合基因疫苗(pcDNA3.1/HisB IL2G1)的免疫效应。方法将pcDNA3.1/HisB IL2G1融合基因疫苗通过肌肉注射途径免疫BALB/c小鼠,每隔3周免疫1次,共免疫3次;酶联免疫法(ELISA)检测抗汉坦病毒(HTNV)抗体;空斑减少中和试验检测免疫血清中和抗体效价;淋巴细胞增殖试验(MTT法)检测免疫小鼠的细胞免疫反应;动物保护试验以BALB/c小鼠为感染动物模型,在第3次免疫后2周,腹腔内注射100TCID50(半数感染量)HTNV,然后以免疫荧光法观察鼠肾切片,评价其保护力。结果pcDNA3.1/HisB IL2G1融合基因疫苗免疫后可刺激机体产生特异性抗体和中和抗体,中和效价为1∶20～1∶80。MTT法结果表明,免疫小鼠的脾淋巴细胞有特异性增殖反应。动物试验表明pcDNA3.1/HisB IL2G1融合基因疫苗能保护小鼠免受HTNV感染。结论pcDNA3.1/HisB IL2G1融合基因疫苗可诱导特异性体液免疫应答和较强的细胞免疫应答。更多还原

【Abstract】 Objective To explore the immune effect of gene vaccine composed of Hantavirus （HTNV） G1 gene and IL-2 gene in BALB/c mice.Methods BALB/c mice were immunized with pcDNA3.1/HisB-IL-2-G1 intramuscularly three times, and 2 boosts were given at 3-week intervals. Serum anti-hantavirus antibodies were detected by ELISA and neutralizing antibodies （NAb） by plaque reduction neutralization test. The lymphocyte proliferation test （MTT assay） was used to test specific cellular immune response to HTNV. BALB/c mice were used to do animal protection experiment. Two weeks after the third immunization, mice were subjected to intraperitoneal challenge with 100 TCID₅0 HTNV, and the protective ability was observed by IFAT. Results The fusion gene vaccine could directly stimulate specific anti-HTNV antibody and neutralizing antibody reaction. The neutralizing antibody titer was 1∶20-1∶80. The specific cellular immune response to HTNV was observed in the immunized mice by lymphocyte proliferation test. Animal experiment revealed that pcDNA3.1/HisB-IL-2-G1 fusion gene could protect the mice from HTNV infection.Conclusion Specific humoral and strong cellular immune responses in mice can be induced with the recombinant eukaryotic expression vector containing Hantavirus G1 gene and IL-2 gene.更多还原