构建两种表达结核分枝杆菌抗原的伊凡诺夫李斯特菌减毒株用于结核病疫苗研究。
Construction of two Listeria ivanovii attenuated strains expressing Mycobacterium tuberculosis antigens for TB vaccine purposes.
作者信息
Lin Qingqing, Zhou Mengying, Xu Zongkai, Khanniche Asma, Shen Hao, Wang Chuan
机构信息
Department of Public Health Laboratory Sciences, Food Safety Monitoring and Risk Assessment Key Laboratory of Sichuan Province, West China School of Public Health, Sichuan University, 16#, Section 3, Ren Min Nan Road, Chengdu, Sichuan 610041, PR China.
Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104-6076, USA; Shanghai Institute of Immunology, Institute of Medical Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai, PR China.
出版信息
J Biotechnol. 2015 Feb 20;196-197:20-6. doi: 10.1016/j.jbiotec.2015.01.008. Epub 2015 Jan 19.
Bacillus Calmette-Guerin (BCG) has failed in complete control of tuberculosis (TB), thus, novel tuberculosis vaccines are urgently needed. We have constructed several TB vaccine candidates, which are characterized by the use of Listeria ivanovii (LI) strain as an antigen delivery vector. Two L. ivanovii attenuated recombinant strains L. ivanovii△actAplcB-Rv0129c and L. ivanovii△actAplcB-Rv3875 were successfully screened. Results from genome PCR and sequencing showed that the Mycobacterium tuberculosis antigen gene cassette coding for Ag85C or ESAT-6 protein respectively had been integrated into LI genome downstream of mpl gene. Western blot confirmed the secretion of Ag85C or ESAT-6 protein from the recombinant LI strains. These two recombinant strains showed similar growth curves as wide type strain in vitro. In vivo, they transiently propagated in mice spleen and liver, and induced specific CD8(+) IFN-γ secretion. Therefore, in this paper, two novel LI attenuated strains expressing specific TB antigens were successfully constructed. The promising growth characteristics in mice immune system and the capability of induction of IFN-γ secretion make them of potential interest for development of TB vaccines.
卡介苗（BCG）未能完全控制结核病（TB），因此，迫切需要新型结核病疫苗。我们构建了几种结核病疫苗候选物，其特征是使用伊凡诺夫李斯特菌（LI）菌株作为抗原递送载体。成功筛选出两种伊凡诺夫李斯特菌减毒重组菌株伊凡诺夫李斯特菌△actAplcB-Rv0129c和伊凡诺夫李斯特菌△actAplcB-Rv3875。基因组PCR和测序结果表明，分别编码Ag85C或ESAT-6蛋白的结核分枝杆菌抗原基因盒已整合到LI基因组中mpl基因的下游。蛋白质免疫印迹法证实了重组LI菌株分泌Ag85C或ESAT-6蛋白。这两种重组菌株在体外显示出与野生型菌株相似的生长曲线。在体内，它们在小鼠脾脏和肝脏中短暂繁殖，并诱导特异性CD8(+) IFN-γ分泌。因此，本文成功构建了两种表达特异性结核抗原的新型LI减毒株。它们在小鼠免疫系统中具有良好的生长特性以及诱导IFN-γ分泌的能力，使其在结核病疫苗开发方面具有潜在的研究价值。
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