Habtamu M, Abrahamsen G, Aseffa A, Andargie E, Ayalew S, Abebe M, Spurkland A
Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, Olso, Norway.
Armauer Hansen Research Institute, Addis Ababa, Ethiopia.
Clin Exp Immunol. 2020 Aug;201(2):187-199. doi: 10.1111/cei.13447. Epub 2020 May 25.
The lack of efficient tools for identifying immunological correlates of tuberculosis (TB) protection or risk of disease progression impedes the development of improved control strategies. To more clearly understand the host response in TB, we recently established an imaging flow cytometer-based in-vitro assay, which assesses multiple aspects of T cell-monocyte interaction. Here, we extended our previous work and characterized communication between T cells and monocytes using clinical samples from individuals with different TB infection status and healthy controls from a TB endemic setting. To identify T cell-monocyte conjugates, peripheral blood mononuclear cells (PBMC) were stimulated with ds-Red-expressing Mycobacterium bovis bacille Calmette-Guérin or 6-kDa early secreted antigenic target (ESAT 6) peptides for 6 h, and analyzed by imaging flow cytometer (IFC). We then enumerated T cell-monocyte conjugates using polarization of T cell receptor (TCR) and F-actin as markers for synapse formation, and nuclear factor kappa B (NF-κB) nuclear translocation in the T cells. We observed a reduced frequency of T cell-monocyte conjugates in cells from patients with active pulmonary tuberculosis (pTB) compared to latent TB-infected (LTBI) and healthy controls. When we monitored NF-κB nuclear translocation in T cells interacting with monocytes, the proportion of responding cells was significantly higher in active pTB compared with LTBI and controls. Overall, these data underscore the need to consider multiple immunological parameters against TB, where IFC could be a valuable tool.
缺乏用于识别结核病（TB）保护的免疫相关因素或疾病进展风险的有效工具，阻碍了改进控制策略的开发。为了更清楚地了解结核病中的宿主反应，我们最近建立了一种基于成像流式细胞仪的体外检测方法，该方法可评估T细胞与单核细胞相互作用的多个方面。在此，我们扩展了之前的工作，并使用来自不同结核病感染状态个体的临床样本以及结核病流行地区的健康对照，对T细胞与单核细胞之间的通讯进行了表征。为了识别T细胞 - 单核细胞结合物，用表达ds - Red的卡介苗或6 kDa早期分泌抗原靶标（ESAT 6）肽刺激外周血单核细胞（PBMC）6小时，然后通过成像流式细胞仪（IFC）进行分析。然后，我们使用T细胞受体（TCR）的极化和F - 肌动蛋白作为突触形成的标志物，以及T细胞中的核因子κB（NF - κB）核转位，来计数T细胞 - 单核细胞结合物。我们观察到，与潜伏性结核感染（LTBI）患者和健康对照相比，活动性肺结核（pTB）患者细胞中T细胞 - 单核细胞结合物的频率降低。当我们监测与单核细胞相互作用的T细胞中的NF - κB核转位时，活动性pTB患者中反应性细胞的比例明显高于LTBI患者和对照。总体而言，这些数据强调了需要考虑针对结核病的多个免疫参数，其中IFC可能是一种有价值的工具。
