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Functional Analysis of the Polymorphic Membrane Protein Family of Chlamydia.

机译：衣原体的多态性膜蛋白家族的功能分析。

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摘要

Chlamydia trachomatis is the most common sexually transmitted bacterial pathogen and the leading cause of infectious blindness. It is an obligate intracellular pathogen that has a biphasic lifecycle encompassing infectious elementary bodies (EBs) and metabolically active reticulate bodies (RBs). C. trachomatis encodes a polymorphic membrane protein (Pmp) gene family whose nine members are differentially expressed under in vitro growth conditions. The observed "phase-variation-like" phenotype of the autotransported Pmps is consistent with observed variable Pmp subtype-specific antibody profiles in patients infected with C. trachomatis. The autotransported Pmps display repeated, often alternating motifs GGA(I,L,V) and FXXN within the predicted "passenger" domain that have been proposed to mediate adhesion of Chlamydia pneumoniae to susceptible cells. The identification of a "magic bullet" adhesin responsible for attachment and subsequent internalization by epithelial cells has eluded researchers for several decades, owing to the inherent difficulties of the Chlamydia system, in particular the genetic intractability of these organisms. Others have established that Pmps of C. pneumoniae and PmpD of C. trachomatis function as adhesins and that PmpD-specific antibodies block attachment. I hypothesize that antibodies against each of the six Pmp subtypes (A, B/C, D, E/F, G/I & H) can neutralize a C. trachomatis infection, and that the application of multiple Pmp subtype-specific antibodies will have a cumulative negative impact on infection. I additionally hypothesize that Pmps of C. trachomatis facilitate adhesion to epithelial cells. For Aim 1, I will investigate the neutralizing effects of single Pmp subtype-specific antibodies by infecting epithelial cells with C. trachomatis serovar E EBs that have been preincubated with antibodies against each of the Pmp subtypes. The inclusions will be visualized using immunofluorescence. For Aim 2, I will investigate Pmp-mediated adherence to epithelial cells using a Pmp-coated latex bead-binding assay. The neutralization assay results cast doubt on the ability of Pmp-specific antibodies to neutralize a chlamydial infection on their own. It is possible that the Pmp-specific antibodies are necessary for complete neutralization, but are not sufficient. Additionally, the adhesion assay results suggest that other functions of Pmps should still be considered, however, their role in adhesion should not be ruled out.
机译：沙眼衣原体是最常见的性传播细菌病原体，也是传染性失明的主要原因。它是专性的细胞内病原体，具有双相生命周期，涵盖传染性基本体（EB）和代谢活性网状体（RB）。沙眼衣原体编码一个多态性膜蛋白（Pmp）基因家族，其九个成员在体外生长条件下差异表达。在沙眼衣原体感染的患者中，观察到的自体转运Pmp的“相变样”表型与观察到的可变Pmp亚型特异性抗体谱一致。自动转运的Pmps在预测的“乘客”域内显示出重复的，经常交替的基序GGA（I，L，V）和FXXN，这些基序已被提议介导肺炎衣原体与易感细胞的粘附。由于衣原体系统固有的困难，特别是这些生物体的遗传难治性，鉴定上皮细胞负责粘附并随后被上皮细胞内在化的“魔术子弹”粘附素已经数十年了。其他人已经证实，肺炎衣原体的Pmps和沙眼衣原体的PmpD起着粘附素的作用，并且PmpD特异性抗体阻断了附着。我假设针对六种Pmp亚型（A，B / C，D，E / F，G / I和H）的抗体可以中和沙眼衣原体感染，并且多种Pmp亚型特异性抗体的应用将对感染有累积的负面影响。我还假设沙眼衣原体的Pmps促进与上皮细胞的粘附。对于目标1，我将通过用已经与每种Pmp亚型的抗体预孵育的沙眼衣原体血清EB感染上皮细胞来研究单个Pmp亚型特异性抗体的中和作用。夹杂物将使用免疫荧光观察。对于目标2，我将使用涂有Pmp的乳胶微珠结合测定法研究Pmp介导的对上皮细胞的粘附。中和测定结果使人怀疑Pmp特异性抗体自行中和衣原体感染的能力。 Pmp特异性抗体可能对于完全中和是必需的，但还不够。此外，粘附测定结果表明，仍应考虑Pmp的其他功能，但是，不应排除它们在粘附中的作用。

著录项

  • 作者

    Grinblat-Huse, Valerie.;

  • 作者单位

    University of Maryland, Baltimore.;

  • 授予单位 University of Maryland, Baltimore.;
  • 学科 Biology Microbiology.;Biology Genetics.
  • 学位 Ph.D.
  • 年度 2014
  • 页码 149 p.
  • 总页数 149
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 地球物理学;
  • 关键词

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