CD4⁺CD8⁺ double-positive T cells are associated with severity of tuberculosis

Tuberculosis (TB) remains a global public health burden, and how immune cell subsets regulate host anti-TB immunity and disease progression remains incompletely understood. While previous studies have focused on single-positive (SP) T cells (CD4⁺ or CD8⁺) in TB pathogenesis, the association between CD4⁺CD8⁺ double-positive (DP) T cells and TB susceptibility, severity, and treatment outcomes have not been fully elucidated. This study aimed to investigate the relationship between DP T cells and other immune cell subsets with TB, and to explore the potential diagnostic and prognostic value of DP T cells in active TB.

A Genome-Wide Association Study (GWAS) was conducted to analyze 731 immune cell traits and a dataset encompassing 895 patients with TB. Subsequently, a cohort including 647 patients with active TB and 632 healthy controls was used to verify the findings of Mendelian randomization (MR). The correlation between the percentage of DP T cells in lymphocytes and TB severity, treatment efficacy, and Mycobacterium tuberculosis (Mtb)-specific IFN-γ production was evaluated. Finally, a random forest model incorporating the percentage of DP T cells in leukocytes and other peripheral blood parameters was constructed to distinguish severe from mild active TB.

MR analysis suggested potential causal links between the percentage of DP T cells among peripheral leukocytes and TB status. Clinical sample validation showed that the percentage of peripheral DP T cell among leukocytes was significantly lower in patients with active TB than in healthy controls (P < 0.001), and was inversely correlated with disease severity. Additionally, the percentage of DP T cells in leukocytes was positively correlated with Mtb-specific antigen-stimulated IFN-γ production. Flow cytometric analysis demonstrated that DP T cells had a significantly higher frequency of IFN-γ-expressing cells compared to CD8⁺ SP T cells (P < 0.001). The constructed random forest model effectively distinguished severe from mild TB, with good diagnostic performance (AUC = 0.985).

Our findings indicate that DP T cells are closely associated with TB severity, and are positively associated with Mtb-specific IFN-γ response. The percentage of peripheral DP T cells in leukocytes could serve as a potential non-invasive biomarker for TB severity stratification.