Monoclonal Antibodies Produced Against Lipopolysaccharide from fimA Type II Porphyromonas gingivalis
Monoclonal Antibodies Produced Against Lipopolysaccharide from fimA Type II Porphyromonas gingivalis
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DOI：
10.1089/hyb.2009.0055
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发表时间：
2009-12-01
期刊：
影响因子：
--
通讯作者：
Abiko, Yoshimitsu
中科院分区：
文献类型：
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作者：
Maruyama, Mitsuhiro;Hayakawa, Mitsuo;Abiko, Yoshimitsu
牙龈卟啉菌是一种重要的牙周病原体，根据菌毛 A (fimA) 的基因型，将其分为六种基因型（I-V 型和 Ib 型）。在这些基因型中，fimA II 型菌株被认为与晚期牙周炎关系最为密切。为了开发被动免疫疗法，利用杂交瘤技术对 fimA II 型牙龈卟啉单胞菌 TDC60 菌株的细胞提取物进行免疫，构建了 300 多个杂交瘤克隆。在这些克隆中，15MAb 通过蛋白质印迹分析识别具有单独梯状结构的 TDC60 脂多糖 (LPS)。对牙龈卟啉单胞菌和大肠杆菌的 TDC60、FDC381（fimA I 型）和 W83（fimA IV 型）的 15MAb 进行进一步的蛋白质印迹分析。这些 MAb 均不能识别大肠杆菌 LPS，并分为至少三种不同的蛋白质印迹模式。为了确认对 LPS 的特异性，选择了三个克隆并使用 TDC60 LPS 进行竞争测定。在以剂量依赖性方式吸收 LPS 后，所有三种 MAb 均降低了针对 TDC60 LPS 的反应性。这些结果表明，成功构建了识别牙龈卟啉单胞菌LPS不同表位的单克隆抗体，这些单克隆抗体可能有助于中和牙龈卟啉单胞菌感染。
An important periodontal pathogen, Porphyromans gingivalis strains are classified into six genotypes (types I-V and Ib), based on the genotype of the fimbriae A (fimA). Among the genotypes, fimA type II strains are thought to be most strongly related to advanced periodontitis. To develop passive immunotherapy, over 300 hybridoma clones were constructed through immunization of cell extracts of fimA type II strain P. gingivalis TDC60 using hybridoma technology. Among these clones, 15MAbs recognized TDC60 lipopolysaccharide (LPS) with an individual ladder-like structure by Western blot analysis. Further Western blotting of the 15MAbs against LPS from TDC60, FDC381 (fimA type I), and W83 (fimA type IV) of P. gingivalis and Escherichia coli was carried out. None of these MAbs recognized E. coli LPS, and divided into at least three different Western blot patterns. To confirm the specificity to LPS, three clones were selected and competition assays were carried out using TDC60 LPS. All three MAbs reduced the reactivity against TDC60 LPS after absorption of the LPS in a dose-dependent manner. These findings suggest that MAbs recognizing different epitopes of P. gingivalis LPS were successfully constructed, and these MAbs may be useful in neutralizing P. gingivalis infection.