Reeves M W, Pine L, Feeley J C, Wells D E
Infect Immun. 1984 Nov;46(2):590-7. doi: 10.1128/iai.46.2.590-597.1984.
Toxic shock toxin (TST), also known as pyrogenic exotoxin C (Schlievert et al., J. Infect. Dis. 143:509-516, 1981) and staphylococcal enterotoxin F (Bergdoll et al., Lancet i:1017-1021, 1981), was purified from toxic shock strains of Staphylococcus aureus by preparative isoelectric focusing and by chromatofocusing. Neither method produced an absolutely pure protein as determined by silver staining of sodium dodecyl sulfate-acrylamide gels, although chromatofocusing was the better method of the two. Three molecular weight variants of the protein were found in the two toxic shock syndrome strains that were studied, regardless of the purification method that was used. An isoelectric point of 7.15 and molecular weights of 21,400, 22,100, and 23,200 were determined for the different forms of the protein from electrophoresis data. A sedimentation coefficient of 2.3S was determined by sucrose gradient centrifugation, and a Stokes radius of 2 X 10(-7) cm was determined by gel filtration. An average molecular weight of 18,900 for all of the TST forms was calculated from these data by the Stokes-Einstein equation. A survey for TST in 32 control and 46 toxic shock strains of S. aureus by isoelectric focusing and by agarose gel double immunodiffusion with specific rabbit antiserum revealed that the isoelectric focusing method tends to overestimate the number of TST-positive strains because of the detection of non-TST, neutral staphylococcal proteins. Based on immunodiffusion data, the association of TST with toxic shock strains was found to be 100% in vaginal isolates and 62% in non-vaginal isolates. In the control strains, TST was found in 16% of the vaginal strains and 23% of the non-vaginal strains. The value of this toxin as a marker for toxic shock and its relationship to the pathogenesis of this disease are discussed.
毒性休克毒素(TST),也被称为致热外毒素C(施利弗特等人,《传染病杂志》143:509 - 516,1981年)和葡萄球菌肠毒素F(伯格多尔等人,《柳叶刀》i:1017 - 1021,1981年),通过制备性等电聚焦和色谱聚焦从金黄色葡萄球菌的毒性休克菌株中纯化得到。通过十二烷基硫酸钠 - 丙烯酰胺凝胶银染测定,两种方法都未产生绝对纯的蛋白质,不过色谱聚焦是两者中较好的方法。在所研究的两株毒性休克综合征菌株中,无论使用何种纯化方法,都发现了该蛋白质的三种分子量变体。根据电泳数据,确定该蛋白质不同形式的等电点为7.15,分子量分别为21,400、22,100和23,200。通过蔗糖梯度离心测定沉降系数为2.3S,通过凝胶过滤测定斯托克斯半径为2×10(-7)厘米。根据这些数据,利用斯托克斯 - 爱因斯坦方程计算出所有TST形式的平均分子量为18,900。通过等电聚焦以及用特异性兔抗血清进行琼脂糖凝胶双向免疫扩散,对32株对照金黄色葡萄球菌菌株和46株毒性休克菌株进行TST检测,结果显示等电聚焦法往往会高估TST阳性菌株的数量,因为会检测到非TST的中性葡萄球菌蛋白质。基于免疫扩散数据,发现TST与毒性休克菌株的关联在阴道分离株中为100%,在非阴道分离株中为62%。在对照菌株中,16%的阴道菌株和23%的非阴道菌株中发现了TST。讨论了这种毒素作为毒性休克标志物的价值及其与该疾病发病机制的关系。