Oliveira Maria Leonor S, Monedero Vicente, Miyaji Eliane N, Leite Luciana C C, Lee Ho Paulo, Pérez-Martínez Gaspar
Centro de Biotecnologia, Instituto Butantan, Av. Vital Brasil 1500, 05503-900, SP, São Paulo, Brazil.
FEMS Microbiol Lett. 2003 Oct 10;227(1):25-31. doi: 10.1016/S0378-1097(03)00645-1.
A number of recent research works in lactic acid bacteria aim towards the design of new strains that could be used as live vectors for the delivery of antigens for oral vaccination, or other therapeutic molecules. In this work, an inducible expression system based on the Lactobacillus casei lactose operon promoter was used to express three important surface antigens of Streptococcus pneumoniae in this lactic acid bacterium: a virulence-related pneumococcal surface antigen (PsaA) and two variants of the virulence factor PspA (pneumococcal surface protein A). Expression of the three proteins was induced upon growth on lactose and strongly repressed by glucose. These proteins were produced intracellularly. Also, secretion to the growth medium was achieved by means of a fusion to the secreting and processing signals from the L. casei surface proteinase. Interestingly, while secreted PspA proteins were found in the culture supernatants, PsaA remained trapped in the cell wall. Expression of pneumococcal antigens in a food-grade organism opens an alternative for mucosal vaccination against this important pathogen.
近期一些关于乳酸菌的研究致力于设计新菌株,这些新菌株可用作活载体,用于口服疫苗抗原或其他治疗分子的递送。在这项研究中,基于干酪乳杆菌乳糖操纵子启动子构建了一个诱导表达系统,用于在该乳酸菌中表达肺炎链球菌的三种重要表面抗原:一种与毒力相关的肺炎球菌表面抗原(PsaA)和毒力因子PspA(肺炎球菌表面蛋白A)的两种变体。这三种蛋白质的表达在乳糖培养基上生长时被诱导,而在葡萄糖存在时则受到强烈抑制。这些蛋白质在细胞内产生。此外,通过与干酪乳杆菌表面蛋白酶的分泌和加工信号融合,实现了向生长培养基的分泌。有趣的是,虽然在培养上清液中发现了分泌型PspA蛋白,但PsaA仍被困在细胞壁中。在食品级生物体中表达肺炎球菌抗原为针对这种重要病原体的黏膜疫苗接种开辟了一条新途径。