DiRienzo J M, Tsai C C, Shenker B J, Taichman N S, Lally E T
Infect Immun. 1985 Jan;47(1):31-6. doi: 10.1128/iai.47.1.31-36.1985.
Hybridoma cell lines which produce monoclonal antibodies to a leukotoxin from Actinobacillus actinomycetemcomitans were prepared. The monoclonal antibodies were selected for their ability to neutralize the cytotoxic activity of the leukotoxin and recognize the toxin on nitrocellulose blots. The antibodies belonged to either the immunoglobulin G1 (IgG1) or IgG2 subclass and differed in their ability to bind to the leukotoxin on nitrocellulose blots. However, only slight differences in neutralization titers were observed. Use of the monoclonal antibodies revealed that polymyxin B-extracted or osmotic shock-released leukotoxin could be separated into several high-molecular-weight polypeptides by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Immunoblot analysis with the monoclonal antibodies also demonstrated that the leukotoxin was present in eight oral strains of A. actinomycetemcomitans that had been previously classified by a biological assay as leukotoxic. The availability of these monoclonal antibodies should facilitate and expand studies concerning the role of the leukotoxin in the pathogenicity of A. actinomycetemcomitans.
制备了产生抗伴放线放线杆菌白细胞毒素单克隆抗体的杂交瘤细胞系。选择这些单克隆抗体是因其具有中和白细胞毒素细胞毒性活性以及在硝酸纤维素印迹上识别该毒素的能力。这些抗体属于免疫球蛋白G1（IgG1）或IgG2亚类，且它们在硝酸纤维素印迹上与白细胞毒素结合的能力有所不同。然而，仅观察到中和效价存在细微差异。使用这些单克隆抗体显示，经多粘菌素B提取或渗透休克释放的白细胞毒素通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳可分离成几种高分子量多肽。用这些单克隆抗体进行的免疫印迹分析还表明，白细胞毒素存在于8株先前通过生物学测定被归类为产白细胞毒素的伴放线放线杆菌口腔菌株中。这些单克隆抗体的可得性应有助于并扩展有关白细胞毒素在伴放线放线杆菌致病性中作用的研究。
