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首页> 外文期刊>Infection and immunity >A macromolecular structure produced by Pseudomonas aeruginosa is recognized by antibody to exotoxin A.
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A macromolecular structure produced by Pseudomonas aeruginosa is recognized by antibody to exotoxin A.

机译：铜绿假单胞菌产生的大分子结构被抗毒素A的抗体识别。

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Organized particulate structures (rods) identified in purified preparations of exotoxin A from culture supernatants of Pseudomonas aeruginosa PA103 were found to be immunochemically cross-reactive with exotoxin A. The rods were visualized by electron microscopy after negative staining as hollow tubes or sheaths (45 by 15 nm). Purified rods were not toxic and not enzymatically active in the ADP-ribosylation assay. Antigenic cross-reactivity between exotoxin A and rods was demonstrated by using monoclonal antibodies directed against either rods or a toxoid of exotoxin A. Hybridoma clones derived from mice immunized with rods or toxoid reacted with both antigens in the enzyme-linked immunosorbent assay. Rods could be dissociated by boiling and resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis into three subunit polypeptides with molecular weights of 70,000, 45,000, and 27,000. Two of the three subunit polypeptides reacted both with antirod and antitoxin monoclonal antibodies after electrophoretic transfer of sodium dodecyl sulfate-polyacrylamide gel electrophoresis-separated proteins to nitrocellulose filters. The results indicate that rods and exotoxin A share common antigenic determinants.
机译：从铜绿假单胞菌PA103的培养上清液中纯化的外毒素A制剂中鉴定出的有组织的颗粒结构（棒）被发现与外毒素A发生免疫化学交叉反应。通过空心镜或鞘进行负染色后，通过电子显微镜观察该棒（45 15 nm）。纯化的棒在ADP-核糖基化测定中无毒且无酶活性。通过使用针对外毒素A的棒或类毒素的单克隆抗体，证明了毒素A和棒之间的抗原交叉反应性。在酶联免疫吸附试验中，来源于用棒或类毒素免疫的小鼠的杂交瘤克隆与两种抗原都发生了反应。棒可以通过煮沸解离，并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分解为三个分子量分别为70,000、45,000和27,000的亚基多肽。将十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离的蛋白质电泳转移到硝酸纤维素滤膜上后，三个亚基多肽中的两个与抗棒和抗毒素单克隆抗体反应。结果表明，棒和外毒素A共有共同的抗原决定簇。

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