Schirmer Sebastian, Rauh Lucas, Alebouyeh Sogol, Delgado-Velandia Mario, Salgueiro Vivian C, Lerma Laura, Serrano-Mestre José L, Azkargorta Mikel, Elortza Félix, Lavín José L, García Maria Jesus, Tórtola Fernández María Teresa, Gola Susanne, Prados-Rosales Rafael
Department of Medical Engineering and Biotechnology, Ernst-Abbe-Hochschule, University of Applied Sciences, Jena, Germany.
Department of Preventive Medicine, Public Health and Microbiology, School of Medicine, Universidad Autónoma de Madrid, Madrid, Spain.
Front Microbiol. 2022 Jun 22;13:907296. doi: 10.3389/fmicb.2022.907296. eCollection 2022.
Tuberculosis (TB) still represents a major global health problem affecting over 10 million people worldwide. The gold-standard procedures for TB diagnosis are culture and nucleic acid amplification techniques. In this context, both lipoarabinomannan (LAM) urine test and rapid molecular tests have been major game changers. However, the low sensitivity of the former and the cost and the prohibitive infrastructure requirements to scale-up in endemic regions of the latter, make the improvement of the TB diagnostic landscape a priority. Most forms of life produce extracellular vesicles (EVs), including bacteria despite differences in bacterial cell envelope architecture. We demonstrated that (), the causative agent of TB, produces EVs and as part of a sophisticated mechanism to manipulate host cellular physiology and to evade the host immune system. In a previous serology study, we showed that the recognition of several mycobacterial extracellular vesicles (MEV) associated proteins could have diagnostic properties. In this study, we pursued to expand the capabilities of MEVs in the context of TB diagnostics by analyzing the composition of MEVs isolated from cultures submitted to iron starvation and, testing their immunogenicity against a new cohort of serum samples derived from TB+ patients, latent TB-infected (LTBI) patients and healthy donors. We found that despite the stringent condition imposed by iron starvation, reduces the number of MEV associated proteins relative to iron sufficient conditions. In addition, TB serology revealed three new MEV antigens with specific biomarker capacity. These results suggest the feasibility of developing a point-of-care (POC) device based on selected MEV-associated proteins.
结核病（TB）仍然是一个重大的全球健康问题，全球有超过1000万人受其影响。结核病诊断的金标准程序是培养和核酸扩增技术。在这种背景下，脂阿拉伯甘露聚糖（LAM）尿液检测和快速分子检测都带来了重大变革。然而，前者灵敏度低，后者成本高且在流行地区扩大规模所需的基础设施要求过高，这使得改善结核病诊断前景成为当务之急。大多数生命形式都会产生细胞外囊泡（EVs），包括细菌，尽管细菌细胞包膜结构存在差异。我们证明，结核病的病原体（）会产生EVs，并且这是其操纵宿主细胞生理和逃避宿主免疫系统的复杂机制的一部分。在之前的一项血清学研究中，我们表明对几种分枝杆菌细胞外囊泡（MEV）相关蛋白的识别可能具有诊断特性。在本研究中，我们通过分析从缺铁培养的菌株中分离出的MEV的组成，并测试它们对一组来自结核病患者、潜伏性结核感染（LTBI）患者和健康供体的新血清样本的免疫原性，来扩大MEV在结核病诊断方面的能力。我们发现，尽管缺铁条件苛刻，但与铁充足条件相比，（）会减少MEV相关蛋白的数量。此外，结核病血清学揭示了三种具有特定生物标志物能力的新MEV抗原。这些结果表明开发一种基于选定的MEV相关蛋白的即时检测（POC）设备是可行的。
