Evaluation of recombinant BCG expressing rotavirus VP6 as an anti-rotavirus vaccine
Evaluation of recombinant BCG expressing rotavirus VP6 as an anti-rotavirus vaccine
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DOI：
10.1016/j.vaccine.2007.01.087
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发表时间：
2007-05-04
期刊：
影响因子：
5.5
通讯作者：
Williamson, Anna-Lise
中科院分区：
文献类型：
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作者：
Dennehy, Maureen;Bourn, William;Williamson, Anna-Lise
在小鼠模型中探索表达轮状病毒VP 6的重组BCG作为抗轮状病毒疫苗。在附加型和整合型大肠杆菌中使用了三个启动子和五个核糖体结合位点。大肠杆菌-分枝杆菌穿梭载体在BCG中表达VP 6。VP 6基因被配置用于在BCG细胞质内积累、从BCG细胞分泌或靶向BCG细胞膜。在稳定性、抗原产生水平、免疫原性和小鼠中的保护方面评估载体。总的不稳定性发生在附加型载体利用热休克蛋白60启动子。然而，三个整合载体使用相同的表达系统和两个附加型载体使用诱导型启动子成功地从BCG回收。前者的增长率并没有令人愉快地降低。后者的生长率大大降低，这意味着存在着显着的代谢负荷。在没有选择的情况下，这些质粒的丢失率很高。VP 6生产水平（占总细胞质蛋白的0.04-1.78%）处于其他rBCG报告范围的下限。一种附加型疫苗和一种整合型疫苗减少了用轮状病毒攻击的腹膜内接种小鼠的病毒脱落。与对照组相比，感染相关的粪便病毒脱落分别减少了66%和62%。这些保护性载体在启动子、核糖体结合位点和抗原产生水平上不同，但都将VP 6蛋白连接到19 kDa脂蛋白信号序列，表明VP 6转运到BCG膜对于诱导保护性免疫应答是重要的。保护发生在血清或粪便中检测不到抗轮状病毒抗体的情况下，这意味着细胞免疫具有保护作用。(C)2007爱思唯尔有限公司保留所有权利。
Recombinant BCG expressing rotavirus VP6 was explored as an anti-rotavirus vaccine in a mouse model. Three promoters and five ribosome-binding sites were used in episomal and integrative E. coli-mycobacterium shuttle vectors to express VP6 in BCG. The VP6 gene was configured for accumulation within the BCG cytoplasm, secretion from the BCG cell or targeting to the BCG cell membrane. Vectors were assessed in terms of stability, levels of antigen production, immunogenicity and protection in mice. Gross instability occurred in episomal vectors utilizing the hsp60 promoter. However, three integrative vectors using the same expression system and two episomal vectors using inducible promoters were successfully recovered from BCG. Growth rates of the former were not delectably reduced. Growth rates of the latter were considerably reduced, implying the existence of a significant metabolic load. In the absence of selection, loss rate of these plasmids was high. VP6 production levels (0.04-1.78% of total cytoplasmic protein) were on the lower end of the range reported for other rBCG. One episomal and one integrated vaccine reduced viral shedding in intraperitoneally vaccinated mice challenged with rotavirus. Compared to controls, infection-associated faecal shedding of virus was reduced by 66% and 62%, respectively. These protective vectors differ in promoter, ribosome-binding site and antigen production level, but both link the VP6 protein to the 19 kDa lipoprotein signal sequence, suggesting that transport of VP6 to the BCG membrane is important for induction of a protective immune response. Protection occurred in the absence of detectable anti-rotavirus antibody in serum or faeces, implicating cellular immunity in protection. (C) 2007 Elsevier Ltd. All rights reserved.