Ren Bing, Szalai Alexander J, Thomas Orlanda, Hollingshead Susan K, Briles David E
Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.
Infect Immun. 2003 Jan;71(1):75-85. doi: 10.1128/IAI.71.1.75-85.2003.
Pneumococcal surface protein A (PspA), a virulence factor of Streptococcus pneumoniae, is exceptionally diverse, being classified into two major families which are over 50% divergent by sequence analysis. A family 1 PspA from strain WU2 was previously shown to impede the clearance of pneumococci from mouse blood and to interfere with complement deposition on the bacterial surface. To determine whether a family 2 PspA can perform the same role as family 1 PspA, the family 1 PspA (from strain WU2) was replaced with a family 2 PspA (from strain TIGR4) by molecular genetic methods to make an isogenic pair of strains expressing different PspA proteins. Surface binding of lactoferrin and interference with C3 deposition by the two types of PspA proteins were determined by flow cytometry, and virulence was assessed in a mouse bacteremia model. Although the family 2 PspA appeared to bind less human lactoferrin than did the family 1 PspA, both PspA proteins could interfere with complement deposition on the pneumococcal surface and could provide full virulence in the mouse infection model. A mutant form of the family 2 PspA with a deletion within the choline-binding region was also produced. Pneumococci with this mutant PspA failed to bind human lactoferrin even though the PspA was present on the pneumococcal surface. The mutant PspA only partially interfered with complement deposition and moderately attenuated virulence. These results suggest that family 1 and family 2 PspA proteins play similar roles in virulence and that surface accessibility of PspA is important for their function.
肺炎球菌表面蛋白A(PspA)是肺炎链球菌的一种毒力因子,具有极高的多样性,通过序列分析可分为两个主要家族,这两个家族的序列差异超过50%。先前研究表明,来自WU2菌株的1型PspA可阻碍肺炎球菌从小鼠血液中清除,并干扰补体在细菌表面的沉积。为了确定2型PspA是否能发挥与1型PspA相同的作用,通过分子遗传学方法将1型PspA(来自WU2菌株)替换为2型PspA(来自TIGR4菌株),从而构建出一对表达不同PspA蛋白的同基因菌株。通过流式细胞术测定两种类型的PspA蛋白与乳铁蛋白的表面结合情况以及对C3沉积的干扰作用,并在小鼠菌血症模型中评估其毒力。尽管2型PspA与1型PspA相比,似乎与人乳铁蛋白的结合较少,但两种PspA蛋白都能干扰补体在肺炎球菌表面的沉积,并且在小鼠感染模型中都具有完全的毒力。还产生了一种2型PspA的突变形式,其胆碱结合区域存在缺失。带有这种突变PspA的肺炎球菌即使PspA存在于肺炎球菌表面,也无法与人乳铁蛋白结合。突变型PspA仅部分干扰补体沉积,并适度减弱毒力。这些结果表明,1型和2型PspA蛋白在毒力方面发挥相似的作用,并且PspA的表面可及性对其功能很重要。