Burns E H, Norman J M, Hatcher M D, Bemis D A
Department of Microbiology, College of Veterinary Medicine, University of Tennessee, Knoxville 37996.
J Clin Microbiol. 1993 Jul;31(7):1838-44. doi: 10.1128/jcm.31.7.1838-1844.1993.
A monoclonal antibody, designated CF8 and prepared against fimbrial protein enrichments of Bordetella bronchiseptica 110H, was determined by immunogold electron microscopy to bind to some but not all fimbrial filaments on intact bacterial cells. Comparison of the reactivity of this antibody with that of monoclonal antibody BPF2, which is specific for Bordetella pertussis serotype 2 fimbriae, indicated that CF8 recognizes an epitope similar to that recognized by BPF2. By Western blot (immunoblot), it was determined that monoclonal antibody CF8 does not react with proteins denatured by treatment with sodium dodecyl sulfate and beta-mercaptoethanol and by boiling for 5 min but that it does recognize fimbrial proteins in their native, nondenatured state. This antibody was used to compare fimbriae between strains of B. bronchiseptica isolated from different species. Strains from pigs, dogs, guinea pigs, and four other species were compared by an enzyme immunoassay. Strains isolated from pigs were found to express significantly more CF8-reactive and B. pertussis serotype 2 cross-reactive fimbriae than strains isolated from guinea pigs. Strains from dogs were more variable in reactivity than those from pigs or guinea pigs. The reactivity with antifimbrial monoclonal antibody CF8 did not correlate with enzyme electromorphotype but did correlate with the host species, suggesting a role for fimbriae in the determination of host species specificity of B. bronchiseptica.
一种名为CF8的单克隆抗体是针对支气管败血波氏杆菌110H的菌毛蛋白富集物制备的，通过免疫金电子显微镜测定，该抗体可与完整细菌细胞上的部分而非全部菌毛丝结合。将此抗体与针对百日咳博德特氏菌2型菌毛的特异性单克隆抗体BPF2的反应性进行比较，结果表明CF8识别的表位与BPF2识别的表位相似。通过蛋白质印迹法（免疫印迹法）测定，单克隆抗体CF8不与经十二烷基硫酸钠和β-巯基乙醇处理并煮沸5分钟而变性的蛋白质发生反应，但能识别天然的、未变性状态的菌毛蛋白。该抗体用于比较从不同物种分离的支气管败血波氏杆菌菌株之间的菌毛。通过酶免疫测定法对来自猪、狗、豚鼠和其他四个物种的菌株进行了比较。结果发现，从猪分离的菌株比从豚鼠分离的菌株表达出显著更多的CF8反应性和百日咳博德特氏菌2型交叉反应性菌毛。狗源菌株的反应性比猪源或豚鼠源菌株更具变异性。与抗菌毛单克隆抗体CF8的反应性与酶电形态型无关，但与宿主物种相关，这表明菌毛在支气管败血波氏杆菌宿主物种特异性的确定中起作用。
