SELECTION OF AN ESCAPE VARIANT OF BORRELIA-BURGDORFERI BY USE OF BACTERICIDAL MONOCLONAL-ANTIBODIES TO OSPB
SELECTION OF AN ESCAPE VARIANT OF BORRELIA-BURGDORFERI BY USE OF BACTERICIDAL MONOCLONAL-ANTIBODIES TO OSPB
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DOI：
10.1128/iai.60.8.3098-3104.1992
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发表时间：
1992-08-01
影响因子：
3.1
通讯作者：
BENACH, JL
中科院分区：
文献类型：
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作者：
COLEMAN, JL;ROGERS, RC;BENACH, JL
两个免疫球蛋白G（IgG）的单克隆抗体（单克隆抗体）的外表面蛋白B（CB 2和CB 6），亲和纯化的小鼠腹水，表现出浓度依赖性的抑制和杀菌特性对伯氏疏螺旋体在螺旋体培养基中孵育24小时后。来自这些单克隆抗体的Fab片段显示出相同的效果，表明它们不是由完整的单克隆抗体凝集生物体引起的。通过分光光度法分析培养基，还检测了含单克隆抗体的培养物中螺旋体生长的抑制。CB 2不抑制赫氏疏螺旋体或B的BEP 4菌株的生长。burgdorferi，两者都不被MAb识别。从杂交瘤上清液中亲和纯化的IgG对B具有类似的作用。结果表明，与腹水IgG一样，其对伯氏螺旋体的抑制和杀菌特性不是由于非特异性有毒污染物。单克隆抗体的杀菌特性不是补体依赖性的，因为在无血清系统中没有。B的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析。暴露于CB 2后存活的伯氏菌揭示了不能表达OspB的逃逸变体。OspA在这些逃逸变体中的持续存在表明OspB的缺乏不是由于含有这两种蛋白质的基因的质粒的丢失。
Two immunoglobulin G (IgG) monoclonal antibodies (MAbs) to outer surface protein B (CB2 and CB6), affinity purified from mouse ascitic fluid, exhibited concentration-dependent inhibitory and bactericidal properties against Borrelia burgdorferi after a 24-h incubation period in spirochete medium. Fab fragments derived from these MAbs showed the same effects, indicating that they were not caused by agglutination of the organisms by the intact MAbs. The inhibition of spirochete growth in cultures containing MAbs was also detected by spectrophotometric analysis of the media. CB2 did not inhibit the growth of Borrelia hermsii or the BEP4 strain of B. burgdorferi, neither of which is recognized by the MAb. Affinity-purified IgG from hybridoma supernatants had similar effects on B. burgdorferi as the ascitic-fluid-derived IgG did, indicating that the inhibitory and bactericidal properties were not due to nonspecific toxic contaminants. The bactericidal properties of the MAbs were not complement dependent as there was none in the serum-free system. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of B. burgdorferi organisms surviving after exposure to CB2 revealed an escape variant which failed to express OspB. The continued presence of OspA in these escape variants indicates that the lack of OspB was not due to the loss of the plasmid which contains the genes for both of these proteins.