论文部分内容阅读
目的:观察巨噬细胞源趋化因子(MDC)基因与柯萨奇病毒B组3型(CVB3)VP1基因融合基因DNA疫苗pcDNA3/MDC-VP1的免疫效果,为CVB3疫苗的研制提供理论和实验依据。方法:6~8周雄性BALB/c纯系小鼠156只随机均分为6组,分别肌内注射0·9%氯化钠溶液(A组)、pcDNA3(B组)、pcDNA3/MDC(C组)、pcDNA3/VP1(D组)、pcDNA3/MDC加pcDNA3/VP1(E组)、pcDNA3/MDC-VP1(F组),3周注射1次,共3次。每次接种后20d断尾取血,测血清中和抗体效价。第3次免疫后3周,每组20只小鼠腹腔注射含10倍半数致死量LD50的CVB3的病毒液各0·2ml,观察各组小鼠的存活情况;每组剩余的6只小鼠腹腔注射含3倍LD50的病毒液各0.2ml,第7天取血后处死,用于检测血中病毒滴度,称量心脏质量计算心脏体重比。结果:A、B、C、D、E和F组的生存率分别为15%、10%、20%、25%、35%和50%,用Kaplan-Meier进行生存分析表明,F组(pcDNA3/MDC-VP1组)的生存状况好于其他各组;F组的血清中和抗体滴度明显提高、血中病毒滴度显著降低,心肌充血肿胀程度较轻。结论:融合基因DNA疫苗pcDNA3/MDC-VP1能诱导小鼠对CVB3VP1产生更强的免疫应答,提高小鼠生存率。
OBJECTIVE: To observe the immunogenicity of DNA vaccine pcDNA3 / MDC-VP1 fused to the VP1 gene of macrophage-derived chemokine (MDC) gene and Coxsackie virus B group 3 (CVB3), and to provide theoretical and experimental evidence for the development of CVB3 vaccine in accordance with. Methods: One hundred and sixty-six male BALB / c mice of 6-8 weeks old were randomly divided into 6 groups: intramuscular injection of 0.9% sodium chloride solution (group A), pcDNA3 (group B) and pcDNA3 / MDC (Group C), pcDNA3 / VP1 (group D), pcDNA3 / MDC plus pcDNA3 / VP1 (group E) and pcDNA3 / MDC-VP1 (group F). 20d after each inoculation bleeding, serum neutralizing antibody titer. Three weeks after the third immunization, 20 mice in each group were intraperitoneally injected with 0.2 ml of CVB3 virus solution containing 10 times the lethal dose of LD50, and the survival of the mice in each group was observed. The remaining 6 mice in each group Intraperitoneal injection of 3 times the LD50 of the virus solution of 0.2ml, the first 7 days after the blood was sacrificed for the detection of blood viral titers, weighed heart mass calculated heart to body weight ratio. Results: The survival rates in groups A, B, C, D, E and F were 15%, 10%, 20%, 25%, 35% and 50%, respectively. Survival analysis using Kaplan- Meier showed that group F (pcDNA3 / MDC-VP1 group) were better than those in other groups. The titer of serum neutralizing antibody in group F was significantly increased, the titer of blood virus was significantly reduced, and the extent of myocardial congestion and swelling was lighter. CONCLUSION: The fusion DNA vaccine pcDNA3 / MDC-VP1 can induce stronger immune response to CVB3VP1 and improve the survival rate of mice.