汉坦病毒H8205株G2-人源IL-2融合基因免疫效果的研究  

Immunization Effect of the Fusion Gene of G2 Segment of Hantavirus H8205 Strain and IL-2 Gene of Human

作　　者：吴志洪[1] 黄汉菊[1] 秦国伟[1] 潘侠[1] 

机构地区：[1]华中科技大学同济医学院基础医学院病原生物学系,武汉430030

出　　处：《华中科技大学学报（医学版）》2005年第1期10-12,16,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目 (No 30170819)

摘　　要：　目的　对比研究两种融合基因pcDNA3 -1/HisB IL2 -G2与pcDNA3. 1/HisB- G2的免疫效果。方法　大量制备重组质粒后免疫Balb/c小鼠, 剂量为100μg/次。于0、4和8周, 共免疫 3 次。免疫前和免疫后第 2、6、10 周采血, 用间接免疫荧光法 (IFA) 与ELISA检测抗汉坦病毒 (HTNV) 抗体; 用空斑减少中和试验 (PRNT) 检测免疫血清的中和效价; 用淋巴细胞增殖试验检测免疫小鼠的细胞免疫反应; 用免疫脾细胞转移保护实验检测疫苗的保护效应。结果　两种融合基因均可刺激机体产生特异性的抗汉坦病毒 76 118 株的交叉抗体和中和抗体, 且差异无显著性意义, 而 pcDNA3 1/HisB- IL2- G2 诱导特异的细胞免疫明显高于 pcDNA3 .1/HisB -G2。结论　pcDNA3. 1/HisB- IL2-G2融合基因的免疫效果优于 pcDNA3 .1/HisB -G2, 该研究结果为进一步研制有效的肾综合征出血热 (HFRS) 基因疫苗提供了重要实验依据。Objective To study the immunization effect of the fusion gene pcDNA3.1/HisB-IL2-G2 in comparison to pcDNA3.1/HisB-G2.Methods The Balb/c mice were immunized three times with two plasmids in 0,4 and 8 weeks.The dose was 100 μg every time.The serum was collected in 0,2,6 and 10 weeks.The anti-HTNV antibodies were detected by IFA,ELISA and the neutralization antibodies were detected by PRNT.The cellular immunity of the immunized mice was examined by T lymphocyte proliferation test (MTT assay).The protective effect of the suckling mice from the hantavirus challenge was detected by a method of passively transferred spleen cells of the immunized mice.Results Both of the two plasmids could directly stimulate specific anti-HTNV 76-118 strain cross-antibody and neutralization antibody reaction,but the difference was not significant.Immunization of the fusion gene pcDNA3.1/HisB-IL2-G2 gained an advantage over that of the fusion gene pcDNA3.1/HisB-G2 in specific cellular immunity.Conclusion The immunization effect of the fusion gene pcDNA3.1/HisB-IL2-G2 is significantly superior to that of the fusion gene pcDNA3.1/HisB-G2,which provides an experimental basis for the further study on the efficient DNA vaccine for hemorrhagic fever with renal syndrome.

关 键 词：融合基因 汉坦病毒 PCDNA3 免疫效果 IL-2 免疫后 细胞免疫反应 重组质粒 保护效应 Balb/c小鼠 

分 类 号：R512.8[医药卫生—内科学] R392[医药卫生—临床医学]