Dept of Immunology, Labor Berlin - Charité Vivantes GmbH, Berlin, Germany.
A. Mantei and T. Meyer contributed equally to this work.
Eur Respir J. 2022 Jul 28;60(1). doi: 10.1183/13993003.01780-2021. Print 2022 Jul.
Rapid and reliable diagnostic work-up of tuberculosis (TB) remains a major healthcare goal. In particular, discrimination of TB infection from TB disease with currently available diagnostic tools is challenging and time consuming. This study aimed at establishing a standardised blood-based assay that rapidly and reliably discriminates TB infection from TB disease based on multiparameter analysis of TB antigen-reactive CD4 T-cells acting as sensors for TB stage-specific immune status.
157 HIV-negative subjects with suspected TB infection or TB disease were recruited from local tertiary care hospitals in Berlin (Germany). Peripheral blood mononuclear cells were analysed for CD4 T-cells reactive to the antigens purified protein derivative and early secretory antigenic target 6 kDa/culture filtrate protein 10. The activation state of TB antigen-reactive T-cells, identified by surface expression of CD154, was evaluated according to the expression profile of proliferation marker Ki-67 and activation markers CD38 and HLA-DR. Using data from 81 subjects with clinically confirmed TB infection (n=34) or culture-proven pulmonary or extrapulmonary TB disease (n=47), 12 parameters were derived from the expression profile and integrated into a scoring system.
Using the scoring system, our assay (TB-Flow Assay) allowed reliable discrimination of TB infection from both pulmonary and extrapulmonary TB disease with high sensitivity (90.9%) and specificity (93.3%) as was confirmed by Monte-Carlo cross-validation.
With low time requirement, ease of sample collection, and high sensitivity and specificity both for pulmonary and extrapulmonary TB disease, we believe this novel standardised TB-Flow Assay will improve the work-up of patients with suspected TB disease, supporting rapid TB diagnosis and facilitating treatment decisions.
快速可靠的结核病（TB）诊断仍然是一个主要的医疗保健目标。特别是，目前可用的诊断工具区分 TB 感染和 TB 疾病具有挑战性且耗时。本研究旨在建立一种标准化的基于血液的检测方法，该方法基于作为 TB 阶段特异性免疫状态传感器的 TB 抗原反应性 CD4 T 细胞的多参数分析，快速可靠地区分 TB 感染和 TB 疾病。
从德国柏林当地的三级保健医院招募了 157 名疑似 TB 感染或 TB 疾病的 HIV 阴性受试者。分析外周血单核细胞对纯化蛋白衍生物和早期分泌抗原靶 6 kDa/培养滤液蛋白 10 的抗原反应性 CD4 T 细胞。根据增殖标志物 Ki-67 和激活标志物 CD38 和 HLA-DR 的表达谱，评估 TB 抗原反应性 T 细胞的激活状态，这些 T 细胞通过 CD154 的表面表达来识别。利用 81 名临床确诊的 TB 感染（n=34）或培养证实的肺或肺外 TB 疾病（n=47）患者的数据，从表达谱中得出 12 个参数并整合到评分系统中。
使用评分系统，我们的检测方法（TB-Flow 检测）允许可靠地区分 TB 感染与肺和肺外 TB 疾病，具有高灵敏度（90.9%）和特异性（93.3%），这一点通过蒙特卡罗交叉验证得到了证实。
由于所需时间短、样本采集简便，以及对肺和肺外 TB 疾病均具有高灵敏度和特异性，我们相信这种新型标准化的 TB-Flow 检测方法将改善疑似 TB 疾病患者的诊断，支持快速 TB 诊断并促进治疗决策。
