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山东大学学报(医学版) ›› 2017, Vol. 55 ›› Issue (7): 38-42.doi: 10.6040/j.issn.1671-7554.0.2016.856

• 基础医学 • 上一篇    下一篇

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

焦凤萍1,张海涛2,赵英会1,田兆菊1,杨树林1   

  1. 1.泰山医学院公共卫生学院临床微生物学教研室, 山东 泰安 271016;2.泰安市妇幼保健院新生儿科, 山东 泰安271016
  • 收稿日期:2016-07-15 出版日期:2017-07-10 发布日期:2017-07-10
  • 通讯作者: 焦凤萍. E-mail:fpjiao@tsmc.edu.cn E-mail:fpjiao@tsmc.edu.cn
  • 基金资助:
    山东省自然科学基金(ZR2013HL057,ZR2013HM033)

Preliminary study of tissue distribution and safety of HSV-2 DNA vaccine in mice

JIAO Fengping1, ZHANG Haitao2, ZHAO Yinghui1, TIAN Zhaoju1, YANG Shulin1   

  1. 1. Department of Clinical Microbiology, School of Public Health, Taishan Medical University, Taian 271016, Shangdong, China;
    2. Department of Neonatology, Maternal and Child Care Service Centre, Taian 271016, Shangdong, China
  • Received:2016-07-15 Online:2017-07-10 Published:2017-07-10

摘要: 目的 初步研究单纯疱疹病毒2型(HSV-2)DNA疫苗pc(pcDNA3.1-)-P6-gBCTL-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-gBCTL-TBK-1肌内注射免疫接种BALB/c小鼠,每只100 μg,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-gBCTL-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。

关键词: 单纯疱疹病毒2型, 安全性, 组织分布, DNA疫苗

Abstract: Objective To investigate preliminarily the tissue distribution and safety of HSV-2 DNA vaccine pc-P6-gBCTL-TBK-1 in mice. Methods The recombinant plasmid pc-P6-gBCTL-TBK-1 was injected into BALB/c mice with 100 μg per mouse for three times at every other week. Heart, liver, spleen, kidney, lung, brain, blood and muscle tissues of immune mice were collected at 24 h, 7 d, 21 d, 35 d and 49 d after inoculation, then total DNAs of tissues were extracted, and the tissue distribution of plasmid DNA and the possibility of integration with host genome were detected by PCR assay. Results At 24 h, 7 d, 21 d and 35 d after the last immunization, the plasmid DNA was detected in heart, liver, spleen, kidney, lung, brain and muscle tissues. At 49 d after the last immunization, the plasmid DNA was detected only in muscle tissue. The gel-purified genomic DNA was detected negative by PCR assay. Conclusion The plasmid pc-P6-gBCTL-TBK-1 can spread quickly to the heart, liver, spleen, kidney, lung, brain, blood and muscle tissues of the mice. With the extention of time, the plasmid DNA only distributes in muscle tissue of vaccination, and the DNA does not integrate with the host genome, which suggests that the plasmid is safe.

Key words: Herpes simplex virus type 2, Tissue distribution, Safety, DNA vaccine

中图分类号: 

  • R373.11
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