Esmael Ahmed, Mihret Adane, Abebe Tamrat, Mussa Daniel, Neway Sebsib, Ernst Joel, Rengarajan Jyothi, Wassie Liya, Howe Rawleigh
Armauer Hansen Research Institute, Addis Ababa, Ethiopia.
Department of Microbiology, Immunology and Parasitology, College of Health Sciences, Addis Ababa University, Ethiopia.
PLoS One. 2025 Sep 3;20(9):e0327243. doi: 10.1371/journal.pone.0327243. eCollection 2025.
Tuberculosis is a major public health challenge in the resource-limited endemic setting of sub-Saharan Africa. The diagnostic challenge becomes worse for smear-negative TB cases. Even if efforts for non-sputum-based TB diagnostic and prognostic biomarkers, there was limited data on blood-based immunological biomarkers among smear-negative PTB patients.Therefore, we assessed the phenotypic profile (HLA-DR, CD-38, Ki-67) of M. tuberculosis specific CD4 + T cells expressing dual IFN-γ and TNF-α cytokines from smear negative PTB patients in Addis Ababa, Ethiopia.
An institutional-based longitudinal cohort study was conducted in Addis Abeba, Ethiopia, on new smear-negative PTB who were adult and HIV-negative in comparison with multiple comparator groups. A total of 149 (confirmed patients with non-TB respiratory disease -33, smear-negative TB-29, smear-positive TB-34, apparently healthy - 53) study participants was enrolled. The expression level of activation (HLA-DR, CD-38) and proliferation (Ki-67) markers from dual IFN-γ and TNF-α cytokines expressing PPD specific CD4 + T cells were assessed after surface and intracellular cytokine staining. To confirm the presence of M. tuberculosis, MGIT/LJ culture, PCR, and smear microscopy were performed.
The overall level of HLA-DR and CD-38 expression in smear-negative and positive pulmonary TB patients were substantially higher than that of confirmed non-TB respiratory illness, apparently healthy QFT positive and negative study participants (p-value = 0.0127, p-value < 0.0001, p-value = 0.0043, p-value <0.0001, respectively) before commencing anti TB treatment. Also, among the smear-negative and positive pulmonary TB cohort, the expression of CD-38, HLA-DR, and HLA-DR + CD-38 + expression was reduced in the second month and six-month cohort compared with baseline data (p-value= < 0.0001, p-value = 0.00365, p -value = 0.0001, respectively).
In this study, we found the diagnostic and prognostic potential of activation markers, particularly CD-38, in smear-negative PTB patients from dual M. tuberculosis-specific IFN-γ + TNF-α+ cytokine producing CD4 + T cells in both the presumed ex vivo and antigen-specific stimulation assays.
在撒哈拉以南非洲资源有限的地方病流行地区，结核病是一项重大的公共卫生挑战。对于涂片阴性的结核病病例，诊断挑战更为严峻。即便人们致力于寻找非痰基结核病诊断和预后生物标志物，但关于涂片阴性肺结核患者血液免疫生物标志物的数据仍然有限。因此，我们评估了来自埃塞俄比亚亚的斯亚贝巴涂片阴性肺结核患者中表达双干扰素-γ和肿瘤坏死因子-α细胞因子的结核分枝杆菌特异性CD4 + T细胞的表型特征（HLA-DR、CD-38、Ki-67）。
在埃塞俄比亚亚的斯亚贝巴开展了一项基于机构的纵向队列研究，研究对象为成年且HIV阴性的新涂片阴性肺结核患者，并与多个对照群体进行比较。总共招募了149名研究参与者（确诊的非结核呼吸道疾病患者-33名、涂片阴性结核病患者-29名、涂片阳性结核病患者-34名、明显健康者-53名）。在进行表面和细胞内细胞因子染色后，评估了表达双干扰素-γ和肿瘤坏死因子-α细胞因子的PPD特异性CD4 + T细胞的激活（HLA-DR、CD-38）和增殖（Ki-67）标志物的表达水平。为确认结核分枝杆菌的存在，进行了MGIT/LJ培养、PCR和涂片显微镜检查。
在开始抗结核治疗前，涂片阴性和阳性肺结核患者中HLA-DR和CD-38的总体表达水平显著高于确诊的非结核呼吸道疾病患者、明显健康的QFT阳性和阴性研究参与者（p值分别为0.0127、p值<0.0001、p值=0.0043、p值<0.0001）。此外，在涂片阴性和阳性肺结核队列中，与基线数据相比，第二个月和第六个月队列中CD-38、HLA-DR以及HLA-DR + CD-38 +的表达均有所降低（p值分别为< 0.0001、p值=0.00365、p值=0.0001）。
在本研究中，我们发现在假定的体外和抗原特异性刺激试验中，激活标志物，特别是CD-38，在来自产生双结核分枝杆菌特异性干扰素-γ +肿瘤坏死因子-α +细胞因子的CD4 + T细胞的涂片阴性肺结核患者中具有诊断和预后潜力。
