Tsai C C, Shenker B J, DiRienzo J M, Malamud D, Taichman N S
Infect Immun. 1984 Feb;43(2):700-5. doi: 10.1128/iai.43.2.700-705.1984.
A leukotoxin from Actinobacillus actinomycetemcomitans was isolated by a procedure that includes polymyxin B extraction, ion-exchange chromatography, and gel filtration chromatography. The procedure resulted in the recovery of 48% of the toxin with a 99-fold increase in specific activity. The isolated toxin has a molecular mass of 180,000 daltons by gel filtration and 115,000 daltons by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It retains all the major biological characteristics previously documented for crude leukotoxin preparations, including susceptibility to heat and proteolytic enzymes and neutralization by sera from patients with juvenile periodontitis. The isolated leukotoxin destroys human but not rat or guinea pig polymorphonuclear leukocytes and has no apparent effect on human erythrocytes. The availability of the A. actinomycetemcomitans leukotoxin should facilitate studies on its chemistry and mode of action as well as its role in the pathogenesis of human periodontal disease.
通过包括多粘菌素B提取、离子交换色谱和凝胶过滤色谱的程序，从伴放线放线杆菌中分离出一种白细胞毒素。该程序回收了48%的毒素，比活性提高了99倍。通过凝胶过滤法测得分离出的毒素分子量为180,000道尔顿，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳测得为115,000道尔顿。它保留了先前粗制白细胞毒素制剂记录的所有主要生物学特性，包括对热和蛋白水解酶的敏感性以及青少年牙周炎患者血清的中和作用。分离出的白细胞毒素可破坏人类多形核白细胞，但不破坏大鼠或豚鼠的多形核白细胞，且对人类红细胞无明显影响。伴放线放线杆菌白细胞毒素的可得性应有助于对其化学性质、作用方式及其在人类牙周疾病发病机制中的作用进行研究。
