Gulig P A, Frisch C F, Hansen E J
Infect Immun. 1983 Nov;42(2):516-24. doi: 10.1128/iai.42.2.516-524.1983.
Six murine plasma cell hybridomas producing monoclonal antibodies (mabs) directed against the 39,000-molecular-weight (39K) major outer membrane protein of Haemophilus influenzae type b were employed in the antigenic analysis of the 39K protein. The initial characterization of the mabs by radioimmunoprecipitation analysis showed that four of these mabs reacted with antigenic determinants of the 39K protein that are exposed on the bacterial cell surface and accessible to antibody. The other two mabs reacted with antigenic determinants of the 39K protein that are either not exposed on the H. influenzae type b cell surface or not accessible to antibody (internal determinants). A total of 126 clinical isolates of H. influenzae type b obtained from pediatric research centers throughout the United States were examined for reactivity with the six mabs by using a solid-phase radioimmunoassay in which bacterial colony growth from agar plates was placed on filter paper and used as antigen. The reactivities of these strains with two of the mabs recognizing cell surface-exposed antigenic determinants of the 39K protein were used to divide the 126 strains into four different groups. Group 1 strains reacted with mab 12D9, group 2 strains reacted with mab 4C4, group 3 strains reacted with both mabs 12D9 and 4C4, and group 4 strains (only one was found) did not react with either mab. The reactivities of two other mabs recognizing cell surface-exposed antigenic determinants of the 39K protein were used to further divide the four groups into eight subgroups. A single mab recognizing an internal antigenic determinant of the 39K protein reacted with every H. influenzae type b strain examined in this study. These data indicate that only limited antigenic heterogeneity exists among the cell surface-exposed antigenic determinants of the 39K outer membrane proteins among H. influenzae type b strains and that at least one internal antigenic determinant of the 39K protein is universally present in all H. influenzae type b strains. Radioimmunoprecipitation analysis also demonstrated that H. influenzae type b strains which lacked a 39K major outer membrane protein possessed a 38K major outer membrane protein which reacted with the anti-39K mabs, indicating that the 38K and 39K outer membrane proteins of different H. influenzae type b strains are antigenically related.
使用六种产生针对b型流感嗜血杆菌39,000分子量（39K）主要外膜蛋白的单克隆抗体（mab）的鼠源浆细胞杂交瘤，对39K蛋白进行抗原分析。通过放射免疫沉淀分析对单克隆抗体进行的初步表征表明，其中四种单克隆抗体与39K蛋白暴露在细菌细胞表面且可被抗体识别的抗原决定簇发生反应。另外两种单克隆抗体与39K蛋白的抗原决定簇发生反应，这些抗原决定簇要么未暴露在b型流感嗜血杆菌细胞表面，要么无法被抗体识别（内部决定簇）。通过固相放射免疫测定法，使用从琼脂平板上生长的细菌菌落放置在滤纸上作为抗原，检测了从美国各地儿科研究中心获得的总共126株b型流感嗜血杆菌临床分离株与这六种单克隆抗体的反应性。这些菌株与两种识别39K蛋白细胞表面暴露抗原决定簇的单克隆抗体的反应性，被用于将126株菌株分为四个不同的组。第1组菌株与单克隆抗体12D9反应，第2组菌株与单克隆抗体4C4反应，第3组菌株与单克隆抗体12D9和4C4都反应，第4组菌株（仅发现一株）与任何一种单克隆抗体都不反应。另外两种识别39K蛋白细胞表面暴露抗原决定簇的单克隆抗体的反应性，被用于将这四个组进一步分为八个亚组。一种识别39K蛋白内部抗原决定簇的单克隆抗体与本研究中检测的每一株b型流感嗜血杆菌菌株都发生反应。这些数据表明，b型流感嗜血杆菌菌株中39K外膜蛋白细胞表面暴露的抗原决定簇之间仅存在有限的抗原异质性，并且39K蛋白的至少一个内部抗原决定簇普遍存在于所有b型流感嗜血杆菌菌株中。放射免疫沉淀分析还表明，缺乏39K主要外膜蛋白的b型流感嗜血杆菌菌株拥有一种与抗39K单克隆抗体发生反应的38K主要外膜蛋白，这表明不同b型流感嗜血杆菌菌株的38K和39K外膜蛋白在抗原上相关。
