Mapping of a protective epitope of the CopB outer membrane protein of Moraxella catarrhalis
Mapping of a protective epitope of the CopB outer membrane protein of Moraxella catarrhalis
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DOI：
10.1128/iai.66.2.540-548.1998
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发表时间：
1998-02-01
影响因子：
3.1
通讯作者：
Hansen, EJ
中科院分区：
文献类型：
--
作者：
Aebi, C;Cope, LD;Hansen, EJ
针对卡他莫拉菌CopB外膜蛋白的单克隆抗体（MAb 10 F3）先前被发现可增强M.在动物模型中的粘膜炎（M.赫尔米宁岛Maciver，J. L.拉蒂默湖D.科普，G. H.小麦克拉肯和E. J.汉森，《感染》。Immun. 61：2003-2010，1993）。在本研究中，显示该相同的MAb在体外针对该病原体发挥补体依赖性杀菌活性。对来自两株MAb 10 F3-反应性和两株MAb 10 F3-非反应性的M.卡他莫拉菌CopB蛋白的氨基酸序列分析表明，这4种CopB蛋白的氨基酸序列至少有90%的同源性。卡他菌O35 E。当在直接酶联免疫吸附测定系统中测试代表这些区域的五种合成肽与MAb 10 F3结合的能力时，显示含有26个氨基酸的寡肽与该MAb结合。通过使用跨越该26聚体的重叠十肽，进一步定位MAb 10 F3的实际结合区域，将含有相同26聚体的融合蛋白与单克隆抗体10 F3结合，免疫小鼠，获得抗血清。在Western印迹分析中，卡他莫拉菌菌株与同源和异源的卡他莫拉菌菌株表面结合。粘膜炎。
A monoclonal antibody (MAb) (MAb 10F3) directed against the CopB outer membrane protein of Moraxella catarrhalis previously was found to enhance pulmonary clearance of M. catarrhalis in an animal model (M. Helminen, I. Maciver, J. L. Latimer, L. D. Cope, G. H. McCracken, Jr., and E. J. Hansen, Infect. Immun. 61:2003-2010, 1993), In the present study, this same MAb was shown to exert complement-dependent bactericidal activity against this pathogen in vitro. Nucleotide sequence analysis of the copB gene from two MAb 10F3-reactive and two MAb 10F3-unreactive strains of M. catarrhalis revealed that the deduced amino acid sequences of these four CopB proteins were at least 90% identical, Comparison of the amino acid sequences of these proteins allowed localization of possible MAb 10F3 binding sites to five relatively small regions of the CopB protein from M. catarrhalis O35E. When five synthetic peptides representing these regions were tested for their ability to bind MAb 10F3 in a direct enzyme-linked immunosorbent assay system, an oligopeptide containing 26 amino acids was shown to bind this MAb, The actual binding region for MAb 10F3 was localized further through the use of overlapping decapeptides that spanned this 26-mer, A fusion protein containing the same 26-mer readily bound MAb 10F3 and was used to immunize mice, The resultant antiserum contained antibodies that reacted with the CopB protein of the homologous M. catarrhalis strain in Western blot analysis and bound to the surface of both homologous and heterologous strains of M. catarrhalis.