摘要
目的观察不同浓度Nogo66蛋白疫苗接种大鼠后产生的免疫效应变化,筛选最佳接种浓度。方法48只SD大鼠随机分为5个实验组和1个对照组,每组8只。实验组大鼠分别接种浓度为50μg/300μl、100μg/300μl、200μg/300μl、400μg/300μl、800μg/300μl的Nogo66蛋白弗氏佐剂疫苗,接种大鼠;对照组不接种疫苗。ELISA法检测不同时间不同浓度组大鼠血清特异性IgG抗体滴度变化,MTT法检测大鼠脾T细胞增殖情况,并进行实验性自身免疫性脑脊髓炎(EAE)评分。结果各实验组大鼠血清IgG抗体滴度从接种第4周开始迅速增高直至第8周,第10周开始达一平台期,此时200μg/300μl组IgG抗体滴度高于其他各浓度组。各实验组大鼠脾T细胞增殖第4周时明显增强,第8周时达最高值;200μg/300μl组从第8周开始活性T细胞水平即高于其他浓度组(P>0.05),于第12周时仍然高于其他浓度组(P<0.05)。所有实验大鼠未发现任何类似EAE的行为变化。结论Nogo66蛋白疫苗接种后,200μg/300μl组IgG抗体滴度和T细胞效应明显强于其他组,且均未发生毒性反应,故200μg/300μl为最佳接种浓度。
Objective To study immunoresponse of SD rats to different dosages of Nogo66 vaccine, and then select the best vaccination concentration. Methods Nogo-66 peptide vaccine plus complete and imcomplete Freunds adjuvant were prepared in concentrations of 50μg/300μl, 100μg/300μl, 200μg/300μl, 400μg/300μl and 800μg/300μl. The vaccines of different concentrations were used to immunize SD rats weekly for 8 weeks, and either complete or imeomplete Freunds adjuvant alone was used as control. Blood was drawn from caudal vein of rats weekly after the first immunization, and the titer of serum special IgG antibody was measured by ELISA. Two animals of every group were sacrificed every four weeks, and the spleens were harvested, and erythroeytes were removed with hypotonic saline. The proliferation rate of the cultured spleen T cells was determined with MTT. Meanwhile the experimental autnimmune encephalomyelitis （EAE） was evaluated for all experimental animals. Results The antibody titer of serum specific IgG of all experimental groups elevated quickly since the 4th week after vaccination, and reached the top value at the 8th week, and it then kept stable for 10 weeks. The IgG antibody titer in 200μg/300μl group was significant higher than that in other experimental groups （ P〈0. 05）. The proliferation of spleen T cells was enhanced obviously at the 4th week, and reached the top value at the 8th week. The proliferation of spleen T cells in 200μg/300μl group was higher than that in other experimental groups after the 8th week, and became considerably higher at the 12th week （P〈0. 05）. Control group had much lower level of IgG antibody titer and the proliferation of the spleen T cells as compared with all the experimental groups during the whole experiment. No EAE was found in the present study. Conclusion After Nogo66 vaccination, the antibody titer is higher and the T cytological effect is stronger in 200μg/300μl group than that in the other experimental groups, therefore 200μg/300μl is suggested as the best vaccination concentration.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2009年第5期630-632,共3页
Medical Journal of Chinese People's Liberation Army
基金
国家自然科学基金资助项目(30572008)
关键词
疫苗
合成
剂量效应关系
免疫
动物实验
vaccines,synthetic
dose-response relationship, immunologic
animal experimentation
