CAJ | 학술논문

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。
목적연구표체광견병독3aG주당단백(GP)적중조복제결함형선병독Ad/GP＇급Ad/GP면역소서후소산생적특이성체액면역응답，체외비세포증식반응，급면역소서대광견병독치사성로내공격적보호력。방법 1×107pfu중조병독경복강대소서진행기출화가강면역，이쾌속형광조억제실험(RFFIT)방법측정소서혈청광견병독특이성중화항체적도，[3H]TdR참입DNA법측정체외비세포증식반응；소서치사성CVS광견병독로내공격측정중조병독면역대소서적보호효과。결과RFFlT방법측정면역소서혈청중화항체적도분별위Ad/GP：0.75 IU/ml，Ad/GP'：2.6 IU/ml；[3H]TdR참입법측정중조병독면역조소서비세포체외수도특이GP항원자격후，증식증강2배이상；86.7％적Ad/GP＇면역소서급66.7％적Ad/GP면역소서가저항약30 LD50CVS광견병독적로내공격。결론표체광견병독G단백적중조복제결함형선병독구유용주기인공정중조병독재체활역묘적호전경。
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.