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编码轴突生长抑制因子的重组DNA疫苗的免疫原性研究

ISSN号：1671-8925
期刊名称：《中华神经医学杂志》
时间：0
分类：Q7[生物学—分子生物学]
作者机构：[1]南方医科大学珠江医院神经外科,广东神经外科研究所,广东脑功能修复与再生重点实验室,广州510282, [2]北京军区总医院神经外科,北京100700
相关基金：国家自然科学基金委员会-广东省人民政府联合基金（U0632008）;广东省科技计划重点项目[粤财教（2008）258-2008A030201019,粤科基办（2007）05/06-7005206]

作者：寇盛斌[1], 姜晓丹[1], 唐艳萍[1], 蔡颖谦[1], 杜谋选[1], 秦玲莎[1], 邹雨汐[1], 徐如祥[2]

关键词：轴突生长抑制因子, DNA疫苗, 免疫原性, 轴突再生, Neurite growth inhibitor, DNA vaccine, Immunogenicity, Axonal regeneration

中文摘要：

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.

英文摘要：

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.

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