小鼠对表达狂犬病毒3aG株糖蛋白的重组复制缺陷型腺病毒的免疫应答

摘要

目的研究表达狂犬病毒3aG株糖蛋白(GP)的重组复制缺陷型腺病毒Ad/GP＇及Ad/GP免疫小鼠后所产生的特异性体液免疫应答，体外脾细胞增殖反应，及免疫小鼠对狂犬病毒致死性颅内攻击的保护力。方法 1×107pfu重组病毒经腹腔对小鼠进行基础和加强免疫，以快速荧光灶抑制实验(RFFIT)方法测定小鼠血清狂犬病毒特异性中和抗体滴度，[3H]TdR掺入DNA法测定体外脾细胞增殖反应；小鼠致死性CVS狂犬病毒颅内攻击测定重组病毒免疫对小鼠的保护效果。结果RFFlT方法测定免疫小鼠血清中和抗体滴度分别为Ad/GP：0.75 IU/ml，Ad/GP'：2.6 IU/ml；[3H]TdR掺入法测定重组病毒免疫组小鼠脾细胞体外受到特异GP抗原刺激后，增殖增强2倍以上；86.7％的Ad/GP＇免疫小鼠及66.7％的Ad/GP免疫小鼠可抵抗约30 LD50CVS狂犬病毒的颅内攻击。结论表达狂犬病毒G蛋白的重组复制缺陷型腺病毒具有用做基因工程重组病毒载体活疫苗的好前景。%Objective To evaluate the immune response of mice immunized with El, E3 deleted replicative deficient recombinant adenovirus which can express glycoprotein of Chinese 3aG rabies virus. Methods Rapid fluorescent focus inhibition test (RFFIT) was used to determine the titer of neutralizing antibodies; specific antigen induced spleen lymphocyte proliferation was determined by in vitro 3H-TdR incorporation assay, and lethal intracerebral challenge with CVS rabies virus was used to evaluate the protective effectiveness of the recombinant virus as candidate vaccine. Results Mice immunized by 1×107 recombinant virus developed high levels of rabies virus neutralizing antibodies (VNA). The spleen lymphocyte of the immunized mice also demonstrated remarkable increased proliferation after stimulation with inactivated rabies virus; 70 %～90 % of the mice immunized by the recombinant virus were protected from lethal intracerebral challenge with CVS rabies virus. Conclusion El, E3 deleted replication defective recombinant adenovirus containing 3aG glycoprotein gene may be developed as effective rabies vaccine to control rabies in China.