PDF(Pubmed)
Abstract：
UNASSIGNED: Shigella is the etiologic agent of a bacillary dysentery known as shigellosis, which causes millions of infections and thousands of deaths worldwide each year due to Shigella\'s unique lifestyle within intestinal epithelial cells. Cell adhesion/invasion assays have been extensively used not only to identify targets mediating host-pathogen interaction, but also to evaluate the ability of Shigella-specific antibodies to reduce virulence. However, these assays are time-consuming and labor-intensive and fail to assess differences at the single-cell level.
UNASSIGNED: Here, we developed a simple, fast and high-content method named visual Adhesion/Invasion Inhibition Assay (vAIA) to measure the ability of anti-Shigellaantibodies to inhibit bacterial adhesion to and invasion of epithelial cells by using the confocal microscope Opera Phenix.
UNASSIGNED: We showed that vAIA performed well with a pooled human serum from subjects challenged with S. sonnei and that a specific anti-IpaD monoclonal antibody effectively reduced bacterial virulence in a dose-dependent manner.
UNASSIGNED: vAIA can therefore inform on the functionality of polyclonal and monoclonal responses thereby supporting the discovery of pathogenicity mechanisms and the development of candidate vaccines and immunotherapies. Lastly, this assay is very versatile and may be easily applied to other Shigella species or serotypes and to different pathogens.
UNASSIGNED: Here, we developed a simple, fast and high-content method named visual Adhesion/Invasion Inhibition Assay (vAIA) to measure the ability of anti-Shigellaantibodies to inhibit bacterial adhesion to and invasion of epithelial cells by using the confocal microscope Opera Phenix.
UNASSIGNED: We showed that vAIA performed well with a pooled human serum from subjects challenged with S. sonnei and that a specific anti-IpaD monoclonal antibody effectively reduced bacterial virulence in a dose-dependent manner.
UNASSIGNED: vAIA can therefore inform on the functionality of polyclonal and monoclonal responses thereby supporting the discovery of pathogenicity mechanisms and the development of candidate vaccines and immunotherapies. Lastly, this assay is very versatile and may be easily applied to other Shigella species or serotypes and to different pathogens.
摘要：
■志贺氏菌是细菌性痢疾的病原体，称为志贺氏菌病，由于志贺氏菌在肠上皮细胞中的独特生活方式，每年在全球范围内导致数百万人感染和数千人死亡。细胞粘附/侵袭试验不仅广泛用于鉴定介导宿主-病原体相互作用的靶标，还要评估志贺氏菌特异性抗体降低毒力的能力。然而,这些试验耗时耗力,无法评估单细胞水平的差异.
■这里，我们开发了一个简单的，一种快速且高含量的方法，称为视觉粘附/侵袭抑制测定（vAIA），通过使用共聚焦显微镜OperaPhenix来测量抗志贺拉抗体抑制细菌粘附和侵袭上皮细胞的能力。
我们显示vAIA与来自用S.sonnei攻击的受试者的合并人血清表现良好,并且特异性抗IpaD单克隆抗体以剂量依赖性方式有效降低细菌毒力。
■vAIA因此可以告知多克隆和单克隆反应的功能，从而支持致病性机制的发现以及候选疫苗和免疫疗法的开发。最后,该测定是非常通用的,并且可以容易地应用于其他志贺氏菌属物种或血清型以及不同的病原体。
■这里，我们开发了一个简单的，一种快速且高含量的方法，称为视觉粘附/侵袭抑制测定（vAIA），通过使用共聚焦显微镜OperaPhenix来测量抗志贺拉抗体抑制细菌粘附和侵袭上皮细胞的能力。
我们显示vAIA与来自用S.sonnei攻击的受试者的合并人血清表现良好,并且特异性抗IpaD单克隆抗体以剂量依赖性方式有效降低细菌毒力。
■vAIA因此可以告知多克隆和单克隆反应的功能，从而支持致病性机制的发现以及候选疫苗和免疫疗法的开发。最后,该测定是非常通用的,并且可以容易地应用于其他志贺氏菌属物种或血清型以及不同的病原体。
