Analysis of the virulence factors of the Streptococcus pyogenes involved in the pathogenesis of toxic shock-like syndrome of Streptococcus pyogenes infection.

化脓性链球菌分泌多种毒素和酶。这些蛋白质以及表面蛋白质被认为是重要的毒力因子。我们提取了中毒性休克样综合征（toxicshock-like syndrome，TSLS）患者化脓性链球菌临床分离株的全部胞外蛋白，并采用双向凝胶电泳技术对其进行了鉴定。同一M血清型临床分离株的蛋白质表达谱基本相同，提示它们引起的临床特征相似。但同一M型的临床分离株，其临床表现不同，蛋白表达谱也不同。通过双向凝胶电泳对分离的蛋白质进行N端氨基酸序列测定，并利用DNA数据库对SpeB、SpeF和链球菌溶血素O进行检测。此外，我们还检测到了几个未报道的蛋白质，我们认为它们可能是毒力因子。我们已经克隆了其中的四个。我们进行了DNA测序，制备了重组蛋白，并发现它们具有DNase活性。我们检测到SpeF，这被认为是在成人呼吸窘迫综合征（ARDS）中发挥重要作用，因为在M1和M3血清型的化脓性链球菌中有相当大的量。但是，我们发现SpeF的N-末端氨基酸不同，等电点也不同。我们现在正在研究不同SpeF点的生物学意义。

Streptococcus pyogenes secretes a variety of toxins and enzymes. These proteins, as well as surface proteins, are considered as important virulence factors. We extracted whole exoproteins of the clinical isolates of S.pyogenes from the patients who suffered toxic shock-like syndrome (TSLS) and characterized them by the two-dimensional gel electrophoresis technique. The protein expression patterns from many of the clinical isolates of the same M serotype were almost same, suggesting that the clinical features caused by them were similar. However some clinical isolates of the same M type which showed different clinical features had different protein expression patterns. We sequenced N-terminal amino acids of the proteins isolated by the two-dimensional gel electrophoresis and detected SpeB, SpeF and streptolysin O by using DNA data bank. In addition, we detected several unreported proteins which we considered as virulence factors. We have already cloned four of them. We performed the DNA sequencing, made recombinant proteins and found that they had DNase activities. We detected SpeF, which is considered to play an important roles in adult respiratory distress syndrome (ARDS), as fairly large amounts in S.pyogenes of both M1 and M3 serotypes. However, we found that SpeF had different N-terminal amino acid and different isoelectricity. We are now studying the biological significance of different SpeF spots.