靶向防龋DNA疫苗在小鼠体内的存留时间分析

Persistence of a Targeted Anti-caries DNA Vaccine in Mice

目的:观察靶向防龋DNA疫苗pG JA-P/VAX免疫小鼠后在免疫原位和引流淋巴结的存留时间。方法:靶向防龋DNA疫苗pG JA-P/VAX分别经肌肉注射和鼻黏膜滴注途径免疫BALB/c小鼠。免疫后1、3、6个月,取肌肉注射组注射局部的肌肉组织和腹股沟引流淋巴结,鼻黏膜滴注组鼻相关淋巴组织和颈部引流淋巴结,提取基因组DNA,以其为模板采用PCR方法扩增pG JA-P/VAX中的GLU序列。结果:免疫后1、3、6个月,在免疫局部组织和引流淋巴结的基因组DNA中均扩增得到870 bp的片段,测序证实该片段序列与GLU序列一致。结论:防龋DNA疫苗pG JA-P/VAX免疫小鼠后,可在免疫原位和引流淋巴结较长期存在。

Objective ： To explore the persistence of a targeted anti - caries DNA vaccine pGJA - P/VAX at both the inoculation sites and the draining lymph nodes. Methods： BALB/c mice were immunized with pGJA -P/VAX by the intramuscular （ i. m. ） or intranasal （ i. n. ） route. 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 cervical lymph nodes from the i.n. immunized groups were collected 1 month, 3 months and 6 months after immunization. The plasmid pGJA - P/VAX at the above sites were detected by PCR. The amplified fragments were analyzed by agarose gel electrophoresis and DNA sequencing. Results： pGJA - P/VAX could be detected at the inoculated muscle tissues, the inguinal lymph nodes, the na- salassociated lymphoid tissues and the draining cervical lymph nodes at all time points were assessed. A 870bp PCR product was amplified. DNA sequening confirmed that the nucleotide sequence of the amplified product was identical with that of the GLU region of pGJA - P/VAX. Conclusion ： pGJA - P/VAX could be detected at both the inoculation sites and the draining lymph nodes for a long period of time after immunization.