Brennan M J, Li Z M, Shahin R D, Burns D L, Nguyen N Y, Liu T Y, Gray M C, Hewlett E L, Manclark C R
Division of Bacterial Products, Food and Drug Administration, Bethesda, MD 20892.
Tokai J Exp Clin Med. 1988;13 Suppl:211-5.
A-69-kDa outer membrane protein present on virulent Bordetella pertussis cells is recognized by the agglutinating monoclonal antibodies BPE3, BPD8, and BPE8. The amino acid composition of this protein, purified from heat extracts of B. pertussis BP353 cells, is different from that of the two major fimbrial antigens of B. pertussis, which is consistent with its being a nonfimbrial protein based on other criteria. Western blot analysis using the monoclonal antibody BPE3 demonstrated that a slightly larger but antigenically cross-reactive protein is also expressed by Bordetella bronchiseptica and Bordetella parapertussis. In addition, a large molecular weight species of about 180-kDa is found in outer membrane extracts of B. bronchiseptica which may represent a precursor form of the protein or indicate that the protein can exist as an oligomer. The monoclonal antibody BPD8 directed against the 69-kDa protein almost completely inhibited the enzymatic activity of adenylate cyclase purified from B. pertussis and also inhibited the intoxication of mammalian cells by this enzyme. Since little enzymatic activity was found associated with the purified 69-kDa protein, these data suggest a role for the 69-kDa protein in regulating the adenylate cyclase toxin of B. pertussis. An additional monoclonal antibody directed against the 69-kDa protein, BPE8, decreases lymphocytosis and delays death in mice receiving a respiratory challenge of virulent B. pertussis cells. These studies suggest that further investigation into the role of this protein as a protective antigen and vaccine candidate is warranted.
存在于有毒百日咳博德特氏菌细胞上的一种69千道尔顿的外膜蛋白可被凝集性单克隆抗体BPE3、BPD8和BPE8识别。从百日咳博德特氏菌BP353细胞的热提取物中纯化得到的这种蛋白质的氨基酸组成，与百日咳博德特氏菌的两种主要菌毛抗原不同，根据其他标准，这表明它是一种非菌毛蛋白。使用单克隆抗体BPE3进行的蛋白质印迹分析表明，支气管败血博德特氏菌和副百日咳博德特氏菌也表达一种稍大但抗原性交叉反应的蛋白质。此外，在支气管败血博德特氏菌的外膜提取物中发现了一种约180千道尔顿的大分子物质，它可能代表该蛋白质的前体形式，或者表明该蛋白质可以以寡聚体形式存在。针对69千道尔顿蛋白质的单克隆抗体BPD8几乎完全抑制了从百日咳博德特氏菌纯化得到的腺苷酸环化酶的酶活性，并且也抑制了该酶对哺乳动物细胞的毒害作用。由于在纯化的69千道尔顿蛋白质中几乎没有发现酶活性，这些数据表明69千道尔顿蛋白质在调节百日咳博德特氏菌的腺苷酸环化酶毒素方面发挥作用。另一种针对69千道尔顿蛋白质的单克隆抗体BPE8，可减少感染有毒百日咳博德特氏菌细胞的小鼠的淋巴细胞增多，并延迟其死亡。这些研究表明，有必要进一步研究这种蛋白质作为保护性抗原和疫苗候选物的作用。
