CAJ | 학술논문

目的探讨以前期实验所构建的腺病毒骨架质粒pAdEasy为载体,编码神经损伤后轴突生长抑制因子Nogo-A、少突胶质细胞髓磷脂糖蛋白(OMgp)、腱糖蛋白-R(TN-R)和髓磷脂相关糖蛋白(MAG)的重组DNA疫苗的免疫原性. 方法 16只5周龄Lewis大鼠按随机数字表法分为DNA疫苗注射组(Vaccine组)和空质粒注射组(pAdEasy组).Vaccine组大鼠以DNA疫苗经双侧胫骨肌注射免疫,1次/周,共持续8周.每次进行疫苗注射前采血和分离血清,Dot-blot和ELISA法对血清中抗体进行定性和定量检测. 结果 6周后Vaccine组大鼠血清能与GST-TN-R和GST-OMgp融合蛋白产生较强的免疫反应,点杂交反应较明显;pAdEasy组大鼠血清则不能与GST-TN-R和GST-OMgp融合蛋白产生免疫反应.6周后Vaccine组大鼠血清中抗体效价则可以达到1:100万,并保持稳定的水平. 结论编码轴突生长抑制因子Nogo-A、OMgp、TN-R和MAG的重组DNA疫苗接种大鼠后能够产生特异性的抗体.说明该重组DNA疫苗具有良好的免疫原性.
목적 탐토이전기실험소구건적선병독골가질립pAdEasy위재체,편마신경손상후축돌생장억제인자Nogo-A、소돌효질세포수린지당단백(OMgp)、건당단백-R(TN-R)화수린지상관당단백(MAG)적중조DNA역묘적면역원성. 방법 16지5주령Lewis대서안수궤수자표법분위DNA역묘주사조(Vaccine조)화공질립주사조(pAdEasy조).Vaccine조대서이DNA역묘경쌍측경골기주사면역,1차/주,공지속8주.매차진행역묘주사전채혈화분리혈청,Dot-blot화ELISA법대혈청중항체진행정성화정량검측. 결과 6주후Vaccine조대서혈청능여GST-TN-R화GST-OMgp융합단백산생교강적면역반응,점잡교반응교명현;pAdEasy조대서혈청칙불능여GST-TN-R화GST-OMgp융합단백산생면역반응.6주후Vaccine조대서혈청중항체효개칙가이체도1:100만,병보지은정적수평. 결론 편마축돌생장억제인자Nogo-A、OMgp、TN-R화MAG적중조DNA역묘접충대서후능구산생특이성적항체.설명해중조DNA역묘구유량호적면역원성.
Objective To detect the immunogenicity of the recombinant DNA vaccine that encoded for neurite growth inhibitors: Nogo-A, oligodendrocyte myelin glycoprotein (OMgp), tenascin-R (TN-R) and myelin-associated glycoprotein (MAG) after the nerve injury under the help of pAdEasy, a kind of adenovirus plasmid being the vector of the DNA. Methods Sixteen 5-w-old Lewis rats were randomized into DNA vaccination group (vaccine group) and pAdEasy group. Rats in the vaccine group were immunized once weekly for a consecutive 8 w by bilateral injection of the recombinant plasmid into the musculus tibialis. The immunized animals in the 2 groups were exsanguinated each time before the vaccination for sera collection, and the qualitation and quantitation of the antibodies in the serum were detected by Dot-blot analysis and ELISA. Results The vaccine group could produce fusion-protein antibodies against Nogo-A, MAG, OMgp and TN-R at the 6th w of vaccine injection, while pAdEasy group could not. The valency of antiserum was shown by ELISA as 1:1 000 000 at the 6th w of vaccine injection and kept this level stably. Conclusion The DNA vaccine exclusively induces the generation of the fusion-protein antibodies against Nogo-A, MAG, OMgp and TN-R in vivo, which controls the favorable immunogenicity.