Centro de Biotecnologia, Instituto Butantan, Av. Vital Brasil 1500, 05503-900 São Paulo, SP, Brazil.
Vaccine. 2009 Dec 9;27(52):7346-51. doi: 10.1016/j.vaccine.2009.09.043. Epub 2009 Sep 25.
Mycobacterium bovis BCG has long been investigated as a candidate for heterologous antigen presentation. We have previously described an rBCG-Pertussis that confers protection against challenge with Bordetella pertussis in neonate and adult mice. In order to obtain stable expression in vivo, we constructed an unmarked BCG lysine auxotrophic and a complementation vector containing the lysine and the genetically detoxified S1 pertussis toxin genes, both under control of the same promoter. Complemented BCG-Delta lysine growth and expression of the pertussis antigen were stable, without the use of an antibiotic marker. Our results show that the complemented rBCG-Delta lysA-S1PT-lysA(+)(kan(-)), which is now suitable to be evaluated in clinical trials, maintains similar characteristics of the original rBCG-pNL71S1PT strain, such as the antigen expression level, cellular immune response and protection against the same model challenge in neonatal-immunized mice.
牛分枝杆菌卡介苗（Mycobacterium bovis BCG）一直被作为异源抗原呈递的候选者进行研究。我们之前曾描述过一种 rBCG-百日咳，它可在新生和成年小鼠中预防百日咳博德特氏菌的挑战。为了在体内获得稳定表达，我们构建了一种非标记的 BCG 赖氨酸营养缺陷型和一个包含赖氨酸和基因解毒的 S1 百日咳毒素基因的互补载体，两者均受相同启动子的控制。补充赖氨酸的 BCG-Delta 生长和百日咳抗原的表达是稳定的，而不使用抗生素标记。我们的结果表明，现在适合在临床试验中评估的互补 rBCG-Delta lysA-S1PT-lysA(+)(kan(-))，保持了与原始 rBCG-pNL71S1PT 菌株相似的特征，例如抗原表达水平、细胞免疫反应和对同一模型挑战的保护作用，在新生免疫小鼠中。
