汉坦病毒包膜糖蛋白G1、G2基因的DNA免疫  

Preliminary study on DNA immunization with hantavirus envelope glycoprotein genes G1 and G2

作　　者：黄玉仙[1] 翁心华[1] 瞿涤[2] 陆群英[3] 朱智勇[3] 

机构地区：[1]复旦大学附属华山医院传染科,上海200040 [2]复旦大学附属华山医院医学分子病毒实验室,上海200040 [3]浙江省疾病控制中心流行性出血热实验室

出　　处：《中华传染病杂志》2004年第3期180-183,共4页Chinese Journal of Infectious Diseases

摘　　要：目的　将编码汉坦病毒浙 37(Z37)株包膜糖蛋白G1、G2的真核表达质粒免疫小鼠 ,观察其能否诱导体液免疫应答。方法　重组质粒 pcDNA3.1 G1、pcDNA3.1 G2以不同方式免疫BALB/C小鼠 ,用免疫荧光测定 (IFA)法检测血特异性抗体 ,用半微量直接免疫酶斑减少中和试验检测血中和抗体。经肌内注射途径免疫 15只BALB/C小鼠 ,3组小鼠分别注射 10 0 μgpcDNA3.1、pcDNA3.1 G1、pcDNA3.1 G2 ;每只小鼠免疫 3次 ,初次免疫后 4周 ,加强 2次 ,间隔 3周。经基因枪免疫 9只BALB/C小鼠 ,3组小鼠分别注射 1.5 μgpcDNA3.1、pcDNA3.1 G1、pcDNA3.1 G2 ;每只小鼠免疫 2次 ,初次免疫后 4周 ,加强 1次。结果　 1.经肌内注射免疫时 ,产生特异性抗体的小鼠个数分别是 :pcDNA3.1 G1组为 5只中 3只 ,pcDNA3.1 G2组为 5只中 2只 ;荧光抗体滴度 1∶2 0～ 1∶80 ;产生中和抗体的小鼠个数分别是 :pcDNA3.1 G1组为 5只中 1只 ,pcDNA3.1 G2组为 5只中 2只 ;中和抗体滴度 1∶10～ 1∶2 0。 2 .经基因枪免疫时 ,实验组小鼠都产生了特异性抗体和中和抗体。荧光抗体滴度 1∶2 0～ 1∶32 0 ,中和抗体滴度≥ 1∶10。结论　重组质粒 pcDNA3.1 G1、pcDNA3.1Objective To examine whether hantavirus Z37 envelope glycoprotein genes G1 and G2 recombinants could elicit humoral immune responses in mice. Methods We vaccinated BALB/C mice with plasmid pcDNA3.1 G1 and pcDNA3.1 G2 respectively by intramuscular and gene gun inoculation. Serum anti hantavirus antibodies were detected by IFA and neutralizing antibodies (NAb) were detected by plaque reduction neutralization test with immunoperoxidase staining. The mice given intramuscular inoculation were divided into 3 groups, each group containing 5 mice: groupⅠas control inoculated with 100 μg pcDNA3.1 ; groupⅡ with 100 μg pcDNA3.1 G1; group Ⅲ with 100 μg pcDNA3.1 G2. Each mouse was immunized three times. 4 weeks after primary vaccination, 2 boosts were given at 3 week intervals. The mice given gene gun inoculation were divided into 3 groups, each group containing 3 mice: groupⅠ as control with 1.5 μg pcDNA3.1; group Ⅱ with 1.5 μg pcDNA3.1 G1; group Ⅲ with 1.5 μg pcDNA3.1 G2.Each mouse was immunized two times. 4 weeks after primary vaccination, 1 boost was given. Results 1. In mice given intramuscular inoculation, serum anti hantavirus antibodies were detected 3 of 5 in group Ⅱ, 2 of 5 in group Ⅲ.IFA titers 1∶20～1∶80. NAb were detected 1 of 5 in group Ⅱ, 2 of 5 in group Ⅲ. NAb titers 1∶10～1∶20. 2.In mice given gene gun inoculation, serum anti hantavirus antibodies and NAb were detected in all experimental group mice, IFA titers 1∶20～1∶320, NAb titers≥1∶10. Conclusions Genetic immunization with HV Z37envelope glycoprotein genes G1 and G2 recombinants could elicit effective humoral immune response in mice.

关 键 词：汉坦病毒属 疫苗DNA 接种 免疫应答 

分 类 号：R392[医药卫生—免疫学]