Vaccines and Immunity Theme, Medical Research Council Unit The Gambia at the London School of Hygiene & Tropical Medicine, Fajara, Gambia.
Department of Clinical Medicine, Institute of Tropical Medicine, Nagasaki University, Nagasaki, Japan.
Front Immunol. 2024 Aug 27;15:1422700. doi: 10.3389/fimmu.2024.1422700. eCollection 2024.
To effectively control tuberculosis (TB), it is crucial to distinguish between active TB disease and latent TB infection (LTBI) to provide appropriate treatment. However, no such tests are currently available. Immune responses associated with active TB and LTBI are dynamic and exhibit distinct patterns. Comparing these differences is crucial for developing new diagnostic methods and understanding the etiology of TB. This study aimed to investigate the relationship between pro- and anti-inflammatory CD4+ cytokine production following stimulation with two types of latency-associated (M.tb) antigens to allow differentiation between active TB and LTBI.
Cryopreserved PBMCs from patients with active TB disease or LTBI were stimulated overnight with replication-related antigen [ESAT-6/CFP-10 (E/C)] or two latency-associated antigens [heparin-binding hemagglutinin (HBHA) and alpha-crystallin-like protein (Acr)]. Responses were analyzed using multiparameter flow cytometry: active TB disease (n=15), LTBI (n=15) and ELISA: active TB disease (n=26) or LTBI (n=27).
CD4+ central memory T cells (Tcm) specific to E/C and CD4+ effector memory T cells specific to Acr and HBHA were higher in LTBI than in TB patients. IFN-γ+Tcm and IL-17+ Tem cells was higher in the LTBI group (p= 0.012 and p=0.029 respectively), but IL-10+ Tcm was higher in the active TB group (p= 0.029) following HBHA stimulation. Additionally, following stimulation with HBHA, IL-10 production from CD4+ T cells was significantly elevated in patients with active TB compared to those with LTBI (= 0.0038), while CD4+ T cell production of IL-17 and IFN-γ was significantly elevated in LTBI compared to active TB (= 0.0076, < 0.0001, respectively). HBHA also induced more CCR6+IL-17+CD4Tcells and IL-17+FoxP3+CD25+CD4Tcells in LTBI than in TB patients (=0.026 and P=0.04, respectively). HBHA also induced higher levels of IFN-γ+IL-10+CD4+ T cells in patients with active TB (Pp=0.03) and higher levels of IFN-γ+IL-17+ CD4+ T cells in those with LTBI (p=0.04). HBHA-specific cytokine production measured using ELISA showed higher levels of IFN-γ in participants with LTBI (P=0.004) and higher levels of IL-10 in those with active TB (P=0.04).
Stimulation with HBHA and measurement of CD4+ T cell production of IFN-γ, IL-10, and IL-17 could potentially differentiate active TB from LTBI. The characteristics of cytokine-expressing cells induced by HBHA also differed between participants with active TB and LTBI.
为了有效控制结核病（TB），区分活动性结核病和潜伏性结核感染（LTBI）以提供适当的治疗至关重要。然而，目前尚无此类检测方法。与活动性 TB 和 LTBI 相关的免疫反应是动态的，并表现出不同的模式。比较这些差异对于开发新的诊断方法和了解 TB 的病因至关重要。本研究旨在探讨两种潜伏相关抗原（M.tb）刺激后促炎和抗炎 CD4+细胞因子产生之间的关系，以区分活动性 TB 和 LTBI。
从活动性 TB 疾病或 LTBI 患者中冷冻保存的 PBMCs 用复制相关抗原[ESAT-6/CFP-10（E/C）]或两种潜伏相关抗原[肝素结合血凝素（HBHA）和α-晶体蛋白样蛋白（Acr）]刺激过夜。使用多参数流式细胞术分析反应：活动性 TB 疾病（n=15），LTBI（n=15）和 ELISA：活动性 TB 疾病（n=26）或 LTBI（n=27）。
与 TB 患者相比，LTBI 中针对 E/C 的 CD4+中央记忆 T 细胞（Tcm）和针对 Acr 和 HBHA 的 CD4+效应记忆 T 细胞更高。在 LTBI 组中，IFN-γ+Tcm 和 IL-17+ Tem 细胞更高（p=0.012 和 p=0.029），但在 HBHA 刺激后，活性 TB 组的 IL-10+Tcm 更高（p=0.029）。此外，与 LTBI 相比，HBHA 刺激后，来自 CD4+T 细胞的 IL-10 产生在活动性 TB 患者中显著升高（=0.0038），而 LTBI 中 CD4+T 细胞产生的 IL-17 和 IFN-γ 在活动性 TB 中显著升高（=0.0076，<0.0001）。与 TB 患者相比，HBHA 还在 LTBI 中诱导更多的 CCR6+IL-17+CD4+T 细胞和 IL-17+FoxP3+CD25+CD4+T 细胞（=0.026 和 P=0.04）。HBHA 还在活动性 TB 患者中诱导更高水平的 IFN-γ+IL-10+CD4+T 细胞（Pp=0.03）和在 LTBI 患者中诱导更高水平的 IFN-γ+IL-17+ CD4+T 细胞（p=0.04）。使用 ELISA 测量的 HBHA 特异性细胞因子产生显示 LTBI 参与者的 IFN-γ 水平更高（P=0.004），而活动性 TB 参与者的 IL-10 水平更高（P=0.04）。
HBHA 刺激和 CD4+T 细胞 IFN-γ、IL-10 和 IL-17 产生的测量可能有助于区分活动性 TB 和 LTBI。HBHA 诱导的细胞因子表达细胞的特征也在活动性 TB 和 LTBI 参与者之间有所不同。
