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目的初步研究单纯疱疹病毒2型(HSV-2)DNA疫苗pc(pc DNA3.1-)-P6-g BCTL-TBK-1在小鼠体内的分布及安全性。方法将前期构建的HSV-2 DNA质粒pc-P6-g BCTL-TBK-1肌内注射免疫接种BALB/c小鼠,每只100μg,分别于第0、2、4周免疫接种3次,于末次免疫后的第24小时及7、21、35、49 d采集小鼠的心、肝、脾、肾、肺、脑、血液和注射部位的肌肉组织,提取各组织DNA,采用PCR法检测质粒DNA在小鼠体内的分布情况及其与宿主基因组发生整合的可能性。结果末次免疫后24 h及7、21、35 d,在小鼠的心、肝、脾、肾、肺、脑、血液和注射部位的肌肉组织中均可检测到质粒,随着时间的延长,在末次免疫后第49天,仅在小鼠注射部位的肌肉组织中能检测到质粒。纯化后的基因组DNA经PCR扩增后均为阴性。结论质粒pc-P6-g BCTL-TBK-1经肌内注射免疫小鼠后,质粒DNA可快速地分布到小鼠的心、肝、脾、肾、肺、脑、血液和肌肉组织中;随着时间的延长,质粒DNA仅分布于接种的肌肉部位,且该DNA疫苗未与宿主的基因组整合,具有一定的安全性。
Objective To study the distribution and safety of HSV-2 DNA vaccine pcDNA3.1 - P6-g BCTL-TBK-1 in mice. Methods BALB / c mice were immunized intraperitoneally with HSV-2 DNA plasmid pc-P6-g BCTL-TBK-1, and each 100 μg was immunized three times at week 0, 2, Twenty-four hours after the last immunization and 7,21,35,49 d, the muscle tissues of the heart, liver, spleen, kidney, lung, brain, blood and the injection site were collected, DNA of each tissue was extracted, and the plasmid The distribution of DNA in mice and the possibility of its integration with the host genome. Results Plasmids were detected in the muscle tissues of heart, liver, spleen, kidney, lung, brain, blood and injection site at 24 h and 7, 21 and 35 d after the last immunization. As time went by, On the 49th day after the last immunization, plasmids were detected only in muscle tissue at the site of injection of the mice. The purified genomic DNA was negative after PCR amplification. Conclusion Plasmid DNA can be quickly distributed to the heart, liver, spleen, kidney, lung, brain, blood and muscle tissues of mice after plasmid pc-P6-g BCTL-TBK-1 was immunized intramuscularly With the extension of time, the plasmid DNA is only distributed in the inoculated muscle site, and the DNA vaccine is not integrated with the host’s genome, with a certain degree of safety.