Verheul A F, Kuipers A J, Braat A K, Dekker H A, Peeters C C, Snippe H, Poolman J T
Eijkman-Winkler Institute of Medical and Clinical Microbiology, Academic Hospital, Utrecht University, The Netherlands.
Clin Diagn Lab Immunol. 1994 Nov;1(6):729-36. doi: 10.1128/cdli.1.6.729-736.1994.
In this study, we characterize the properties of nine monoclonal antibodies (MAbs) that recognize meningococcal lipopolysaccharides (LPS). The following three specific MAbs that had not been described previously were elicited in BALB/c mice by using an immunotype L3,7,9 oligosaccharide-tetanus toxoid conjugate in combination with Quil A: 4D1-B3, 3A12-E1, and 4A8-B2. These MAbs reacted with L3,7,9 LPS on immunoblots and in the LPS enzyme-linked immunosorbent assay (ELISA) and recognised strains containing L3, L3,7, L8 (except 3A12-E1), or L9 LPS in the whole-cell ELISA. The six other MAbs have been described in previous studies (K. Saukkonen, M. Leinonen, H. Abdillahi, and J.T. Poolman, Vaccine 7:325-328, 1989; R.J.P.M. Scholten, B. Kuipers, H.A. Valkenburg, J. Danjert, W.D. Zollinger, and J.T. Poolman, J. Med. Microbiol., in press) and were obtained after immunization with outer membrane protein complexes containing LPS: MN15A11, MN15A8-1, MN15A17-1, MN11A11G, MN14F20-11, and MN14F21-11. MN15A11 was specific for L3,7,9 LPS and displayed properties similar to those of 3A12-E1. MN15A17-1, MN14F20-1, and MN11A11G were cross-reactive, and MN14F21-11 was specific for the L1,8 immunotype. Epitope specificities of MAbs reacting with L3,7,(8),9 strains were analyzed. MAbs 4D1-B3, 3A12-E1, and 4A8-B2 recognized phosphoethanolamine group-containing oligosaccharide-specific epitopes. MN15A11 and MN15A17-1 were probably directed against a conformational epitope, although for MN5A11 recognition of an unknown L3,7,9-specific epitope in the 2-keto-3-deoxyoctulosonic acid (KDO)-lipid A region cannot be excluded. MN15A8-1, a strongly cross-reactive MAb, recognized a determinant which included the KDO-lipid A region and the more terminal saccharides.(ABSTRACT TRUNCATED AT 250 WORDS)
在本研究中,我们对九种识别脑膜炎球菌脂多糖(LPS)的单克隆抗体(MAb)的特性进行了表征。通过使用免疫型L3,7,9寡糖 - 破伤风类毒素缀合物与Quil A联合,在BALB/c小鼠中诱导产生了以下三种先前未描述过的特异性单克隆抗体:4D1 - B3、3A12 - E1和4A8 - B2。这些单克隆抗体在免疫印迹和LPS酶联免疫吸附测定(ELISA)中与L3,7,9 LPS发生反应,并在全细胞ELISA中识别含有L3、L3,7、L8(3A12 - E1除外)或L9 LPS的菌株。其他六种单克隆抗体已在先前的研究中有所描述(K. Saukkonen、M. Leinonen、H. Abdillahi和J.T. Poolman,《疫苗》7:325 - 328,1989;R.J.P.M. Scholten、B. Kuipers、H.A. Valkenburg、J. Danjert、W.D. Zollinger和J.T. Poolman,《医学微生物学杂志》,即将发表),它们是在用含有LPS的外膜蛋白复合物免疫后获得的:MN15A11、MN15A8 - 1、MN15A17 - 1、MN11A11G、MN14F20 - 11和MN14F21 - 11。MN15A11对L3,7,9 LPS具有特异性,并且表现出与3A12 - E1相似的特性。MN15A17 - 1、MN14F20 - 1和MN11A11G具有交叉反应性,而MN14F21 - 11对L1,8免疫型具有特异性。分析了与L3,7,(8),9菌株反应的单克隆抗体的表位特异性。单克隆抗体4D1 - B3、3A12 - E1和4A8 - B2识别含磷酸乙醇胺基团寡糖特异性表位。MN15A11和MN15A17 - 1可能针对构象表位,尽管对于MN5A11而言,不能排除其识别2 - 酮 - 3 - 脱氧辛酸(KDO) - 脂质A区域中未知的L3,7,9特异性表位。MN15A8 - 1是一种强交叉反应性单克隆抗体,识别一个包括KDO - 脂质A区域和更末端糖类的决定簇。(摘要截断于250字)