Schlievert P M, Kelly J A
Ann Intern Med. 1982 Jun;96(6 Pt 2):982-6. doi: 10.7326/0003-4819-96-6-982.
Staphylococcal pyrogenic exotoxin type C was purified by differential precipitation with ethanol and resolubilization in water followed by thin-layer isoelectric focusing. The purified toxin migrated as a homogeneous protein when re-electrofocused in polyacrylamide (isoelectric point, 7.2), when subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (molecular weight, 22,000 daltons), and when reacted against hyperimmune antisera in an Ouchterlony immunodiffusion plate. A 2200-fold purification of the toxin could be obtained. Pyrogenic exotoxin C was elaborated by all of 44 Staphylococcus aureus isolates from patients with toxic shock syndrome when the strains were tested in a blind manner, but five of 37 control isolates produced toxin (p less than 0.001). Furthermore, all of 91 additional isolates from patients with toxic shock syndrome from diverse geographic locations were positive for toxin. In contrast, 158 of 609 (26%) S. aureus isolates not associated with toxic shock syndrome and one of 22 Staphylococcus epidermidis isolates were positive for pyrogenic exotoxin type C. The biological properties of pyrogenic exotoxin C included pyrogenicity, enhancement of susceptibility to endotoxin shock, enhancement of skin reactivity to give a positive Dick reaction, suppressions of IgM synthesis, and nonspecific T-lymphocyte mitogenicity.
通过乙醇分级沉淀、水溶再溶解及薄层等电聚焦法纯化葡萄球菌C型致热外毒素。纯化后的毒素在聚丙烯酰胺中再次进行等电聚焦时(等电点为7.2)、进行十二烷基硫酸钠聚丙烯酰胺凝胶电泳时(分子量为22,000道尔顿)以及在Ouchterlony免疫扩散平板中与超免疫抗血清反应时,均呈现为单一的蛋白质迁移带。该毒素可实现2200倍的纯化。当对来自中毒性休克综合征患者的44株金黄色葡萄球菌菌株进行盲法检测时,所有菌株均能产生致热外毒素C,但37株对照菌株中有5株产生毒素(p<0.001)。此外,从不同地理位置的中毒性休克综合征患者中分离出的另外91株菌株均产毒素阳性。相比之下,609株与中毒性休克综合征无关的金黄色葡萄球菌菌株中有158株(26%)以及22株表皮葡萄球菌菌株中有1株产葡萄球菌C型致热外毒素阳性。葡萄球菌C型致热外毒素的生物学特性包括致热性、增强对内毒素休克的易感性、增强皮肤反应性使狄克反应呈阳性、抑制IgM合成以及非特异性T淋巴细胞促有丝分裂性。