Table_1_Monitoring Anti-tuberculosis Treatment Response Using Analysis of Whole Blood Mycobacterium tuberculosis Specific T Cell Activation and Functional Markers.DOCX

BackgroundBlood-based biomarkers have been proposed as an alternative to current sputum-based treatment monitoring methods in active tuberculosis (ATB). The aim of this study was to validate previously described phenotypic, activation, and cytokine markers of treatment response in a West African cohort.
MethodsWhole blood immune responses to Mycobacterium tuberculosis ESAT-6/CFP-10 (EC) and purified protein derivative (PPD) were measured in twenty adults at baseline and after 2 months of standard TB treatment. Patients were classified as fast or slow responders based on a negative or positive sputum culture result at 2 months, respectively. Cellular expression of activation markers (CD38, HLA-DR), memory markers (CD27), and functional intracellular cytokine and proliferation (IFN-γ, Ki-67, TNF-α) markers were measured using multi-color flow cytometry.
ResultsThere was a significant increase in the proportion of CD4+CD27+ cells expressing CD38 and HLA-DR following EC stimulation at 2 months compared to baseline (p = 0.0328 and p = 0.0400, respectively). Following PPD stimulation, slow treatment responders had a significantly higher proportion of CD8+CD27–IFN-γ+ (p = 0.0105) and CD4+CD27+HLA-DR+CD38+ (p = 0.0077) T cells than fast responders at baseline. Receiver operating curve analysis of these subsets resulted in 80% sensitivity and 70 and 100% specificity, respectively (AUC of 0.82, p = 0.0156 and 0.84, p = 0.0102).
ConclusionOur pilot data show reductions in expression of T cell activation markers were seen with treatment, but this was not associated with fast or slow sputum conversion at 2 months. However, baseline proportions of activated T cell subsets are potentially predictive of the subsequent speed of response to treatment.

研究背景：血液生物标志物（blood-based biomarkers）已被提出作为当前活动性肺结核（active tuberculosis, ATB）患者基于痰液的治疗监测方法的替代方案。本研究旨在在西非队列中验证此前已报道的与治疗应答相关的表型、活化及细胞因子标志物。
研究方法：本研究纳入20名成人受试者，于基线及标准抗结核治疗2个月后，检测其全血对结核分枝杆菌（Mycobacterium tuberculosis）ESAT-6/CFP-10（EC）与结核菌素纯蛋白衍生物（purified protein derivative, PPD）的免疫应答反应。根据受试者治疗2个月时的痰液培养结果阴/阳性，分别将其归类为快速应答者与慢速应答者。采用多色流式细胞术检测活化标志物（CD38、HLA-DR）、记忆标志物（CD27）以及功能性胞内细胞因子与增殖标志物（IFN-γ、Ki-67、TNF-α）的细胞表达水平。
研究结果：与基线相比，经EC刺激后，治疗2个月时表达CD38与HLA-DR的CD4+CD27+细胞比例显著升高（分别为p=0.0328与p=0.0400）。经PPD刺激后，慢速应答者在基线时的CD8+CD27–IFN-γ+与CD4+CD27+HLA-DR+CD38+ T细胞比例均显著高于快速应答者（分别为p=0.0105与p=0.0077）。对上述细胞亚群进行受试者工作特征曲线（Receiver Operating Characteristic, ROC）分析显示，其灵敏度为80%，特异性分别为70%与100%（曲线下面积（Area Under the Curve, AUC）分别为0.82，p=0.0156；0.84，p=0.0102）。
研究结论：本初步研究数据显示，抗结核治疗后T细胞活化标志物的表达水平有所降低，但该变化与治疗2个月时的痰液阴转快慢并无关联。然而，基线时活化T细胞亚群的比例可潜在预测后续治疗应答的速度。