摘要
目的分析鼠伤寒沙门菌(St)细菌影负载防龋DNA疫苗对黏膜免疫效能的影响。方法收集St减毒株J357,同时转入噬菌体Phi X基因E表达质粒与pREP4质粒,加入异丙基-β-D-硫代半乳糖苷(IPTG)进行诱导,收集负载防龋DNA疫苗。按照随机原则,将20只SPF级BALB/c雌性小鼠分为A组(布比卡因包裹p VXA1 5μg)、B组(布比卡因包裹p GJGLU/VAX 5μg)、C组(细菌影负载空载体p VXA1 5μg)、D组(细菌影负载防龋DNA疫苗p GJGLU/VAX 5μg)共4组经鼻免疫小鼠,每组均为5只。采用酶联免疫吸附试验对各组小鼠唾液抗体的产生情况进行检测。结果经IPTG诱导后,对照组细菌生长速度较快;而表达基因E经IPTG诱导后,大量细菌出现死亡,随着诱导时间的延长,细菌的死亡数量明显进一步增多。经IPTG诱导前,取携带基因E的St中J357活菌数为2×1012个/L;而经异丙基-β-D-硫代半乳糖苷(IPTG)诱导后,其活菌数仅为5×108个/L,表明大量细菌被杀死。经酶联免疫吸附试验检测结果发现,唾液总Ig A抗体水平在各组小鼠免疫前的比较,并无显著差异(P> 0. 05);相比A组,C、D组小鼠免疫后唾液总Ig A抗体水平均明显升高(P <0. 05);而C、D组唾液总Ig A抗体水平与B组比较,均无显著差异(P> 0. 05)。经酶联免疫吸附试验检测结果发现,各组小鼠免疫前均未检出唾液特异性抗葡聚糖结合区Ig A抗体;免疫后8周,D组唾液特异性抗葡聚糖结合区Ig A抗体水平较A、B、C组均明显升高(P <0. 05),而B组抗体水平与A、C组比较,均无显著差异(P> 0. 05)。结论经鼻黏膜途径免疫小鼠经St细菌影负载防龋DNA疫苗后可明显改善其免疫效能。
Objective To analyze the effect of Salmonella typhimurium(St)bacterial ghost-loaded anti-caries DNA vaccine on mucosal immunity.Methods The attenuated St strain J357 was collected and transfected into E expression plasmid and pREP4 plasmid of PhiX gene.Induced by IPTG,DNA vaccine loaded with caries prevention was collected.According to the random principle,20 SPF BALB/c female mice were divided into four groups:group A(bupivacaine encapsulated pVXA1 5μg),group B(bupivacaine encapsulated pGJGLU/VAX 5μg),group C(bacterial ghost-loaded empty vector pVXA1 5μg),and group D(bacterial ghost-loaded anti-caries DNA vaccine pGJGLU/VAX 5μg),with 5 mice in each group.The production of St in mice of each group was detected by enzyme linked immunosorbent assay.Results After IPTG induction,the growth rate of bacteria in the control group was faster,while after IPTG induction,a large number of bacteria died.With the prolongation of induction time,the number of bacterial death increased significantly.Before induction by IPTG,the number of viable St J357 carrying gene E was 2×1012/L,while after induction by IPTG,the number of viable St J357 was only 5×108/L,indicating that a large number of bacteria were killed.The results of enzyme linked immunosorbent assay showed that there was no significant difference in the total IgA antibody level in saliva between groups before immunization(P>0.05).Compared with group A,the total IgA antibody level in saliva of group C and D increased significantly(P<0.05),while the total IgA antibody level in saliva of group C and D had no significant difference compared with group B(P>0.05).The results of enzyme linked immunosorbent assay showed that no salivary specific anti-glucan binding IgA antibody was detected in all groups before immunization.Eight weeks after immunization,the level of salivary specific anti-glucan binding IgA antibody in group D was significantly higher than that in group A,B and C(P<0.05),but there was no significant difference between group B and group A and C(P>0.05).Con
作者
马琼
李玲
孙聪
邓春妮
马丽
MA Qiong;LI Ling;SUN Cong(Department of Stomatology,the First Affiliated Hospital of Xi'an Jiaotong University,Xi'an Shaanxi 710061,China.)
出处
《临床和实验医学杂志》
2019年第6期584-587,共4页
Journal of Clinical and Experimental Medicine
基金
陕西省软科学研究计划项目(编号:2016KRM127)
关键词
小鼠
鼠伤寒沙门菌
细菌影
龋齿
DNA疫苗
Mice
Salmonella typhimurium
Dacterial ghost
Dental caries
DNA vaccine
