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  

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

作　　者：焦凤萍[1] 张海涛[2] 赵英会[1] 田兆菊[1] 杨树林[1] JIAO Fengping ZHANG Haitao ZHAO Yinghui TIAN Zhaoju YANG Shulin(Department of Clinical Microbiology, School of Public Health, Taishan Medical University, Taian 271016, Shangdong, China Department of Neonatology, Maternal and Child Care Service Centre, Taian 271016, Shangdong, China)

机构地区：[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

出　　处：《山东大学学报（医学版）》2017年第7期38-42,共5页Journal of Shandong University:Health Sciences

基　　金：山东省自然科学基金(ZR2013HL057;ZR2013HM033)

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。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 Ixg per mouse for three times at every other week. Heart, liver, spleen, kidney, lung, brain, blood and mus- cle 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 de- tected 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 mus- cle 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.

关 键 词：DNA疫苗 单纯疱疹病毒2型 组织分布 安全性 

分 类 号：R373.11[医药卫生—病原生物学]