USE OF THE NIRB PROMOTER TO DIRECT THE STABLE EXPRESSION OF HETEROLOGOUS ANTIGENS IN SALMONELLA ORAL VACCINE STRAINS - DEVELOPMENT OF A SINGLE-DOSE ORAL TETANUS VACCINE
USE OF THE NIRB PROMOTER TO DIRECT THE STABLE EXPRESSION OF HETEROLOGOUS ANTIGENS IN SALMONELLA ORAL VACCINE STRAINS - DEVELOPMENT OF A SINGLE-DOSE ORAL TETANUS VACCINE
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DOI：
10.1038/nbt0892-888
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发表时间：
1992-08-01
期刊：
影响因子：
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通讯作者：
FAIRWEATHER, NF
中科院分区：
文献类型：
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作者：
CHATFIELD, SN;CHARLES, IG;FAIRWEATHER, NF
将质粒pTETnir15导入鼠伤寒沙门氏菌aroA aroD活口服疫苗菌株BRD 509中，该质粒指导破伤风毒素的无毒免疫原性片段C从厌氧诱导型nirB启动子表达。将所得菌株（命名为BRD 847）用于口服接种BALB/c小鼠，并测试其质粒稳定性及其抵抗致死性破伤风毒素攻击的能力。pTETnir 15通过在免疫小鼠的组织中生长或存留的细菌稳定地遗传，而另一种BRD 509衍生物，命名为BRD 753，携带指导来自tac启动子的片段C表达的质粒pTET 85，是高度不稳定的。用单次口服剂量的BRD 847免疫的小鼠产生了高水平的循环抗片段C抗体，并且对破伤风毒素攻击具有牢固的保护作用。用单次口服剂量的BRD 753免疫的小鼠未产生可检测的抗片段C抗体。加强免疫后，检测到抗体，但小鼠仅部分免受破伤风毒素攻击。因此，使用体内诱导型启动子如nirB可以是获得沙门氏菌疫苗株中异源抗原的稳定体内表达的普遍适用的方法。
Plasmid pTETnir15, which directs the expression of the non-toxic immunogenic fragment C of tetanus toxin from the anaerobically inducible nirB promoter, was introduced into the Salmonella typhimurium aroA aroD live oral vaccine strain BRD509. The resulting strain, designated BRD847, was used to vaccinate orally BALB/c mice and was tested for plasmid stability and its ability to protect against a lethal tetanus toxin challenge. pTETnir15 was stably inherited by bacteria growing or persisting in the tissues of immunized mice whereas another BRD509 derivative, designated BRD753, harboring plasmid pTET85 which directs fragment C expression from the tac promoter, was highly unstable. Mice immunized with a single oral dose of BRD847 developed high levels of circulating anti-fragment C antibodies and were solidly protected against tetanus toxin challenge. Mice immunized with a single oral dose of BRD753 developed no detectable anti-fragment C antibodies. After boosting, antibodies were detected, but the mice were only partially protected against tetanus toxin challenge. Thus the use of an in vivo inducible promoter such as nirB may be a generally applicable approach to obtaining the stable in vivo expression of heterologous antigens in Salmonella vaccine strains.