Hsieh Wen-Shyang, Yang Yi-Yuan, Yang Hsin-Yi, Huang Yu-Shan, Wu Hsueh-Hsia
Department of Laboratory Medicine, Taipei Medical University-Shuang Ho Hospital, New Taipei City, Taiwan; School of Medical Laboratory Science and Biotechnology, Taipei Medical University, Taipei, Taiwan; Graduate Institute of Biomedical Informatics, Taipei Medical University, Taipei, Taiwan.
School of Medical Laboratory Science and Biotechnology, Taipei Medical University, Taipei, Taiwan.
J Microbiol Immunol Infect. 2016 Jun;49(3):329-34. doi: 10.1016/j.jmii.2014.07.012. Epub 2014 Oct 8.
Although the mortality rates have decreased over the past few decades, neonatal meningitis is still a severe disease with high morbidity. Moreover, approximately 40% of survivors exhibit neurological sequelae. Escherichia coli is the major Gram-negative bacterial pathogen in neonatal meningitis. The N-terminal β-barrel domain of the outer membrane protein A (OmpA) of E. coli is essential for effective protein conformation and function and contains four surface-exposed hydrophilic loops. In this study, we expressed different fragments of the four ring structures of the N-terminal domain, and investigated whether these recombinant OmpA fragments can protect mice from death after E. coli infection.
We expressed the recombinant proteins of the following OmpA fragments by using molecular cloning of Loop 1-2, Loop 1-3, Loop 1-4, Loop 2-3, Loop 2-4, and Loop 3-4. Animal experiments were subsequently performed to investigate the effects of these recombinant OmpA fragments on the survival of C57BL/6 mice after intracerebral E. coli RS218 administration.
This study demonstrated that the recombinant Loop 1-3, Loop 2-3, and Loop 2-4 fragments of OmpA can protect mice from intracerebral E. coli infection.
In bacterial meningitis, although antibiotic therapy is the first choice for management, neurological complications can seldom be averted. Based on the results of the present study, we intend to establish an effective therapeutic application for E. coli meningitis.
尽管在过去几十年中死亡率有所下降，但新生儿脑膜炎仍然是一种发病率很高的严重疾病。此外，约40%的幸存者存在神经后遗症。大肠杆菌是新生儿脑膜炎中的主要革兰氏阴性菌病原体。大肠杆菌外膜蛋白A（OmpA）的N端β桶结构域对于有效的蛋白质构象和功能至关重要，并且包含四个表面暴露的亲水性环。在本研究中，我们表达了N端结构域四个环结构的不同片段，并研究了这些重组OmpA片段是否能保护小鼠免受大肠杆菌感染后的死亡。
我们通过对环1-2、环1-3、环1-4、环2-3、环2-4和环3-4进行分子克隆来表达以下OmpA片段的重组蛋白。随后进行动物实验，以研究这些重组OmpA片段对脑内注射大肠杆菌RS218后C57BL/6小鼠存活的影响。
本研究表明，OmpA的重组环1-3、环2-3和环2-4片段可保护小鼠免受脑内大肠杆菌感染。
在细菌性脑膜炎中，尽管抗生素治疗是首要治疗选择，但神经并发症很少能避免。基于本研究结果，我们打算为大肠杆菌脑膜炎建立一种有效的治疗应用方法。
