【作者】 许庆安； 樊明文； 边专； 彭彬； 贾荣；

【Author】 XU Qing-an, FAN Ming-wen, BIAN Zhuan, PENG Bin, JIA Rong.(School of Stomatology. Wuhan University, Wuhan 430079,Hubei Province, China)

【机构】 武汉大学口腔医学院牙体牙髓科,口腔生物医学工程教育部重点实验室；

【摘要】 目的:观察靶向防龋质粒pGJA-P／VAX免疫小鼠后在免疫局部及引流淋巴结的存留时间。方法:将Bupivacaine-pGJA-P／ VAX复合体和Bupivacaine-pVAX1复合体分别经肌内注射途径和鼻黏膜滴注途径免疫BALB／c小鼠,免疫后1、3、6个月采集免疫部位的肌肉组织、腹股沟引流淋巴结、鼻相关淋巴组织和颈部引流淋巴结。从以上组织中提取基因组,PCR方法扩增pGJA-P／ VAX的GLU片段,琼脂糖凝胶电泳检测扩增片段大小,DNA序列测定核苷酸序列。结果:免疫后1、3、6个月,肌内注射组免疫部位的肌肉组织和腹股沟引流淋巴结、鼻黏膜滴注组的鼻相关淋巴组织和颈部引流淋巴结中均扩增获得870bD的PCR扩增片断, DNA序列测定证实与GLU片断核苷酸序列一致。结论:靶向防龋质粒pGJA-P／VAX免疫小鼠后可在免疫局部及引流淋巴结中较长时间存留,提示免疫局部组织和引流淋巴结可作为质粒储存库不断产生抗原物质,刺激机体的免疫系统维持免疫应答。更多还原

【Abstract】 PURPOSE: To observe the persistence of the targeted anti-caries DNA plasmid pGJA-PA AX at the inoculation sites and the draining lymph nodes after immunization of mice. METHODS: BALB/c mice were immunized with Bupivacaine: pGJA-P/ VAX complexe or Bupivacaine:pVAX1 complexe intramuscularly or intranasally. Mice were sacrificed at 1 month, 3 month and 6 month after the initial immunization. Muscle tissues at the inoculation sites and the draining inguinal lymph nodes from the i.m. immunized groups and the nasal-associated lymphoid tissues and the draining cenical lymph nodes from the i.n. immunized groups were then collected. Plasmids at the above sites were detected by PCR. The amplified fragments were analyzed by agarose gel electrophoresis and DNA sequencing. RESULTS: PCR analysis showed that pGJA-P/VAX could be detected at the inoculated muscle tissues, the inguinal lymph nodes, nasal-associated lymphoid tissues and the draining cervical lymph nodes at all time points assessed. A 870bp PCR product was amplified. DNA sequencing confirmed that the nucleotide sequence of the amplified product was identical with that of pGJA-P/VAX. No fragment was amplified in the pVAX1 immunized mice. CONCLUSION: The persistent immune responses generated after pGJA-P/VAX immunization are likely owing to the DNA depot in the organism, which acts as a booster immunization.更多还原