Abstract
OBJECTIVES: The early diagnosis and immunoregulatory mechanisms of active tuberculosis (ATB) and latent tuberculosis infection (LTBI) remain unclear, and the role of metabolic genes in host-pathogen interactions requires further investigation. METHODS: Single-cell RNA sequencing (scRNA-seq) was applied to analyze peripheral blood mononuclear cells (PBMCs) from 7 individuals, including 2 healthy controls (HC), 2 LTBI patients, and 3 ATB patients. We identified T-cell-associated metabolic differentially expressed genes (TCM-DEGs) through integrated differential expression analysis and machine learning algorithms (XGBoost, SVM-RFE, and Boruta). These TCM-DEGs were then used to construct a diagnostic model and evaluate its clinical applicability. RESULTS: The analysis revealed significant immunological alterations in TB patients, characterized by markedly elevated monocyte/macrophage populations (p < 0.001) accompanied by reduced T and NK cell counts. Notably, LTBI cases demonstrated an intermediate CD4+/CD8+ T-cell ratio, indicative of dynamic immune homeostasis. The TB cohort exhibited increased inflammatory T-cell populations, while CD8+ T-cell-mediated MHC-I and BTLA signaling pathways were identified as key regulators of immune clearance and modulation. Transcriptomic profiling identified five metabolically significant differentially expressed genes (FHIT, MAN1C1, SLC4C7, NT5E, AKR1C3; p < 0.05) that effectively distinguish between latent tuberculosis infection (LTBI) and active tuberculosis (TB). The machine learning-driven diagnostic framework demonstrated remarkable consistency across independent validation cohorts (GSE39940, GSE39939), exhibiting AUC values spanning 0.867-0.873. Molecular subtyping analysis delineated two distinct TB phenotypes: an immune-activated M1 macrophage-dominant subtype and a CD8 + T-cell infiltrated immunophenotype. Clinical validation substantiated the differential expression patterns of T-cell-related metabolic differentially expressed genes (TCM-DEGs; p < 0.05), while the nomogram predictive model achieved exceptional discriminative capacity (C-index = 0.944), demonstrating superior clinical applicability through decision curve analysis. CONCLUSIONS: Our findings reveal that TCM-DEGs critically regulate TB progression through immune-metabolic reprogramming and cell-cell communication networks. The developed diagnostic model and molecular subtyping strategy enable precise TB-LTBI differentiation and inform immunotherapy optimization.