摘要
目的比较小鼠巨噬细胞源趋化因子(macrophage-derived chemokine,MDC)和志贺毒素B亚单位(Shiga tox-in B subunit,STxB)与柯萨奇病毒B组3型(coxsackievirus B3,CVB3)VP1融合DNA疫苗的免疫效果。方法将120只雄性BALB/c小鼠分为6组,每组20只,分别肌内注射pcDNA3、pcDNA3/MDC、pcDNA3/STxB、pcDNA3/VP1、pcDNA3/MDC-VP1和pcDNA3/STxB-VP1,每3周1次,共3次,每次免疫后20d取血清,用微量中和试验检测CVB3中和抗体,第3次免疫后3周,每组取3只小鼠,取脾脏制备脾细胞,用CCK-8法检测特异性CTL杀伤活性,其余小鼠用10LD50的CVB3攻击,观察小鼠的生存情况。结果pcDNA3/VP1、pcDNA3/STxB-VP1和pcDNA3/MDC-VP1组均能诱导小鼠产生中和抗体;除pcDNA3/STxB-VP1组外,另两组抗体滴度随免疫次数增加而提高;第3次免疫后,pcDNA3/MDC-VP1组平均抗体滴度和脾淋巴细胞特异性CTL杀伤活性高于pcDNA3/VP1(P<0.05)。病毒攻击后,pcDNA3/MDC-VP1组21d生存率高于其他各组(P<0.05)。pcDNA3/STxB-VP1组抗体滴度和生存情况与pcDNA3/VP1组无明显差异。结论融合基因疫苗pcDNA3/MDC-VP1能诱导小鼠产生较强的体液和细胞免疫,提高了小鼠生存率,免疫效果优于pcDNA3/STxB-VP1。
Objective To compare the effect of macrophage-derived chemokine （MDC） and Shiga toxin B subunit （STxB） gene on the neutralizing antibodies and specific CTL response induced by Coxasckievirus B3 （CVB3） VP1 fusion DNA vaccine, as wells as the protective effect of the genes on the mice challenged with CVB3. Methods Male BALB/c mice were divided into 6 groups randomly, 20 for each, and injected imtramuscularly （ i. m. ） pcDNA3, pcDNA3/MDC, pcDNA3/STxB, pcDNA3/VP1, pcDNA3/MDC-VP1, pcDNA3/STxB-VP1, once for 3 weeks, totally for 3 times. Serum samples 20 d after each injection were taken and detected for the neutralizing antibody against CVB3 by micro-neutralization test. Three weeks after the third immunization, splenocytes from 3 mice of each group were isolated to detect specific CTL response by CCK-8 assay. The mice were challenged with 10LD50 of CVB3 after the 3 injections. The survival time and survival rate were observed. Results Neutralizing antibodies were induced in pcDNA3/VP1, pcDNA3/STxB-VP1 and pcDNA3/MDC-VP1 groups, and the antibody titers were increased with the increasing number of injections except pcDNA3/STxB-VP1 group. After the third inoculation, the mean neutralizing antibody titers in pcDNA3/ MDC-VP1 group were higher than pcDNA3/VP1 and pcDNA3/STxB-VP1 groups （P 〈 0.05）. Up to the 21st day after CVB3 challenge, the survival rates of pcDNA3/MDC-VP1 group were better than others （ P 〈 0.05）. However, neutralizing antibody and survival rate had no significant changes in pcDNA3/VP1 and pcDNA3/ STxB-VP1 groups. Conclusion pcDNA3/MDC-VP1 vaccine can induce a stronger immunological effect, result in a higher survival rate, and had better influence than pcDNA3/STxB-VP1.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2008年第9期799-802,共4页
Journal of Third Military Medical University
基金
河北省自然科学基金(C2004000631)
河北省邢台市2006年科学研究与发展指导计划项目(2006SP098)~~
