单纯疱疹病毒2型DNA疫苗在小鼠体内分布及安全性的初步分析

发布时间：2019-05-16 14:58

【摘要】：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。
[Abstract]:Objective to study the distribution and safety of herpes simplex virus type 2 (HSV-2) DNA vaccine pc (pc DNA3.1- P6 g BCTL-TBK-1 in mice. Methods BALB/c mice were immunized with HSV-2 DNA plasmid pc-P6-g BCTL-TBK-1 intramuscular at 100 渭 g each for 3 times in 0, 2 and 4 weeks, respectively. The muscle tissues of heart, liver, spleen, kidney, lung, brain, blood and injection site were collected at 24 hours and 7, 21, 35 and 49 days after the last immunization, and DNA, was extracted from each tissue. The distribution of plasmid DNA in mice and the possibility of integration with host genome were detected by PCR. Results plasmid was detected in heart, liver, spleen, kidney, lung, brain, blood and muscle tissue of mice 24 h and 7 h, 21 d, 35 d after the last immunization. With the prolongation of time, on the 49th day after the last immunization, the plasmid could be detected in the muscle tissue of the mouse heart, liver, spleen, kidney, lung, brain, blood and injection site. The plasmid could be detected only in the muscle tissue of the injection site of mice. The purified genomic DNA was negative after PCR amplification. Conclusion after intramuscular injection of plasmid pc-P6-g BCTL-TBK-1 into mice, plasmid DNA can be rapidly distributed in heart, liver, spleen, kidney, lung, brain, blood and muscle tissue of mice. With the prolongation of time, plasmid DNA was only distributed in the inoculated muscle, and the DNA vaccine was not integrated with the host genome, so it was safe to some extent.
【作者单位】：泰山医学院公共卫生学院临床微生物学教研室;泰安市妇幼保健院新生儿科;
【基金】：山东省自然科学基金(ZR2013HL057,ZR2013HM033)
【分类号】：R392

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