Almeida MR, van Hooij A, Pierneef L
… +12 more, Wassenaar G, da Silva Corrêa R, de Almeida Ribeiro RS, Dos Santos LKC, de Oliveira AL, Santos AP, Mafort TT, Rufino R, de Macedo CS, Corstjens PLAM, Rodrigues LS, Geluk A
Despite significant efforts to improve diagnostics, tuberculosis (TB) diagnosis still relies on clinical and laboratory methods limited by cost, expertise, and time. Early diagnosis is essential for effective management...Despite significant efforts to improve diagnostics, tuberculosis (TB) diagnosis still relies on clinical and laboratory methods limited by cost, expertise, and time. Early diagnosis is essential for effective management of pulmonary TB (PTB), pleural TB (PLTB), and latent TB infection (LTBI), highlighting the urgent need for rapid, accurate, and accessible TB diagnostics based on low-invasive sampling. In this study, a quantitative rapid test, previously evaluated in European and African cohorts, was applied to a Brazilian cohort to measure six host-derived proteins in serum from individuals with PTB, PLTB, other pleural diseases (OPLD), LTBI, and controls. Serum levels of CRP, ferritin, and SAA1/A2 were significantly elevated in PTB and PLTB compared to controls. SAA1/A2 and IP-10 levels were higher in PLTB compared to LTBI. Furthermore, IP-10 serum levels showed diagnostic potential, as they were elevated in PTB compared with both LTBI and controls, and significantly increased in PLTB compared to OPLD. In pleural effusion, IP-10, IL-6, and S100A12 were significantly higher in PLTB than in OPLD, while ferritin levels were higher in OPLD. Our study shows that quantitative detection of host-derived biomarkers identified for active PTB in European and African populations can support clinical decision-making for TB spectrum in a Brazilian setting.
BACKGROUND: Tuberculous meningitis (TBM), the most severe form of tuberculosis, may involve the gut-brain axis. This study investigated the link between TBM, brain white matter integrity measured by diffusion tensor imag...BACKGROUND: Tuberculous meningitis (TBM), the most severe form of tuberculosis, may involve the gut-brain axis. This study investigated the link between TBM, brain white matter integrity measured by diffusion tensor imaging (DTI), and the gut microbiome. METHODS: 22 TBM patients and 31 healthy controls underwent MRI and provided fecal samples. Gut microbiota (16S rRNA sequencing), fecal metabolites (metabolomics), and DTI metrics (FA, MD, RD, AD) were analyzed. Tract-based spatial statistics and correlation analyses were employed. RESULTS: TBM patients showed lower gut microbiota α-diversity. The abundance of the Escherichia genus correlated negatively with FA and positively with MD in specific white matter tracts. Metabolomics revealed elevated acetic acid in TBM patients, which correlated with both Escherichia abundance and DTI metrics. KEGG analysis indicated altered arginine and proline metabolism pathways. CONCLUSION: TBM is associated with differences in gut microbiota composition. Higher relative abundance of Escherichia is linked to white matter microstructural damage, potentially mediated by specific bacterial metabolites.
Inflammatory microenvironments regulate immune cell differentiation, activation, and programming. Although traditionally considered terminally differentiated effectors, neutrophils can acquire distinct functional states...Inflammatory microenvironments regulate immune cell differentiation, activation, and programming. Although traditionally considered terminally differentiated effectors, neutrophils can acquire distinct functional states in response to environmental cues. To examine context-dependent neutrophil programming, cells from healthy donors were cultured in vitro under cytokine regimens mimicking opposing inflammatory environments (GM-CSF/IFN-γ or IL-4/IL-13/TGF-β), alone or combined with PMA and Mycobacterium tuberculosis, to assess functional and transcriptional responses. GM-CSF/IFN-γ-conditioned neutrophils exhibited increased cell size, altered nuclear morphology, enhanced MHC class II and CD86 expression, and elevated IL-8 and reactive oxygen species production. This profile was associated with increased TNF-α, IL-10, TLR2, and TLR4 gene expression and reduced global DNA methylation. In contrast, IL-4/IL-13/TGF-β-conditioned neutrophils resembled non-conditioned controls regarding surface markers and cytokine production but showed reduced ROS generation and increased DNMT3A expression. Upon in vitro infection with M. tuberculosis, GM-CSF/IFN-γ-conditioned neutrophils produced higher IL-8 and IL-1β levels and formed more neutrophil extracellular traps. Additionally, plasma from patients with severe tuberculosis modulated TLR4, CCR7, and IP-10 expression in healthy neutrophils, indicating systemic inflammatory influences. These findings demonstrate that inflammatory conditioning induces coordinated transcriptional and epigenetic remodeling, shaping neutrophil antimicrobial and immunoregulatory potential in infectious contexts.
BACKGROUND: The Bacille Calmette-Guérin (BCG) vaccine against tuberculosis is the most widespread vaccine in the world. Discovered by French investigators Albert Calmette and Camille Guérin at the Pasteur Institute, it r...BACKGROUND: The Bacille Calmette-Guérin (BCG) vaccine against tuberculosis is the most widespread vaccine in the world. Discovered by French investigators Albert Calmette and Camille Guérin at the Pasteur Institute, it remains the only effective vaccine against tuberculosis infection. This report describes the recognition and identification of a previously unknown French handwritten laboratory notebook prepared by Drs. Calmette and Camille Guérin recording their experiments performed during the development of the BCG vaccine. METHODS: The notebook was examined, translated into English, photographed and the experiments analyzed. RESULTS: The manuscript laboratory notebook consists of 69 leaves written in 2 hands, one of which corresponds to that of Albert Calmette. It contains details of experiments that were performed during the development of the BCG vaccine at the Pasteur Institute by Drs. Calmette and Guérin. These include experimental inoculations of rabbits and guinea pigs; descriptions of the pathology of skin lesions, inflammatory reactions, and organ pathology; and survival outcome. The experiments describe varying inoculative dosages of the bacteria, and different routes of administration including intraperitoneal and subcutaneous injections, and administration of bacilli in the ear. In those cases where the animal had died following inoculation of tubercle bacilli, necropsy was performed and the organs examined and the pathology findings described. Details of culture experiments and vaccine passage are listed. CONCLUSIONS: This previously unknown notebook is a highly organized and detailed record of investigations using tuberculosis in animal experiments and microbiological culture to produce a safe and effective vaccine, first used in humans in 1921.
Pulmonary and skin infections caused by Mycobacterium abscessus, a rapidly growing non-tuberculous mycobacterium (NTM), are becoming increasingly prevalent worldwide. Managing these infections is particularly challenging...Pulmonary and skin infections caused by Mycobacterium abscessus, a rapidly growing non-tuberculous mycobacterium (NTM), are becoming increasingly prevalent worldwide. Managing these infections is particularly challenging due to the bacterium's inherent resistance to many antibiotics, often resulting in extended treatment periods or therapeutic failure. With few effective drugs available, identifying new therapeutic targets is critical. A promising approach involves targeting the pathogen's energy metabolism, specifically the electron transport chain (ETC), which plays a vital role in its survival and virulence. This review explores the essential components of the ETC in Mycobacterium abscessus as potential drug targets to support and expedite the development of novel treatments.
OBJECTIVE: The aim of the study was to determine the associations between vitamin D (VD) and cytokine levels in active pulmonary tuberculosis (PTB) patients with chronic pulmonary aspergillosis (CPA). METHODS: One hundre...OBJECTIVE: The aim of the study was to determine the associations between vitamin D (VD) and cytokine levels in active pulmonary tuberculosis (PTB) patients with chronic pulmonary aspergillosis (CPA). METHODS: One hundred seventy-two active PTB patients, twenty-eight PTB patients with CPA (PTB-CPA), and sixty healthy subjects were included in the study. The concentrations of serum 25-hydroxyvitamin D, interleukin (IL)-1β, IL-4, IL-6, IL-8, IL-10 and tumor necrosis factor (TNF-α) were measured by ELISA. RESULTS: A significantly lower VD level was detected in PTB-CPA patients (14.96 ± 10.89 ng/mL) than in both PTB patients (20.72 ± 8.07 ng/mL, p < 0.01) and controls (23.90 ± 8.16 ng/mL, p < 0.001). VD deficiency was significantly more prevalent in PTB-CPA patients than in PTB patients (OR = 2.86, 95% CI: 1.22-6.69; p = 0.01) or controls (OR = 5.33, 95% CI: 2.02-14.11; p = 0.0005). In the PTB group, VD levels exhibited a significant inverse correlation with only IL-6 (r = -0.376, p = 0.0001), whereas in PTB-CPA patients, stronger inverse correlations were observed with IL-6 (r = -0.691, p = 0.0005) and IL-8 (r = -0.538, p = 0.01). In cavitary PTB patients, a significant inverse link was found between VD and IL-1β and IL-6 levels (all p ≤ 0.05). CONCLUSION: Our data revealed a high prevalence of VD deficiency in all groups, predominantly in PTB-CPA patients. The inverse correlations between VD levels and IL-6 and IL-8 levels in PTB-CPA patients as well as between VD and IL-1β and IL-6 levels in cavitary PTB patients, provide further evidence for the role of VD as a regulator of inflammatory pathways in the context of coinfection.
A significant proportion of individuals heavily exposed to infectious tuberculosis patients do not acquire Mycobacterium tuberculosis (Mtb) infection, as detected by an interferon gamma release assay (IGRA). Trained immu...A significant proportion of individuals heavily exposed to infectious tuberculosis patients do not acquire Mycobacterium tuberculosis (Mtb) infection, as detected by an interferon gamma release assay (IGRA). Trained immunity may contribute to this host resistance to Mtb infection, also termed early clearance. From a prospective tuberculosis household study in Indonesia we selected 80 heavily exposed IGRA-negative household contacts, of whom 40 converted their baseline-negative IGRA to positive after three months (IGRA converters) and 40 remained IGRA-negative (early clearers). From all individuals we measured circulating β-D-glucan and used their serum for induction of trained immunity in vitro, using peripheral blood mononuclear cells from healthy unexposed Dutch donors, that were stimulated with unrelated stimuli six days after exposure to serum from household contacts. β-D-glucan concentrations and positivity did not correlate with early clearance, nor with serum-induced in-vitro trained immunity as measured by heterologous cytokine responses. These findings suggest that early clearance is unlikely to be maintained by circulating β-D-glucan or other serum factors present at exposure to Mtb, and may instead depend on cell-intrinsic or local host processes not captured by serum-based assays.
BACKGROUND: Type 2 diabetes mellitus elevates the risk of developing tuberculosis, posing a substantial challenge to TB treatment and control. However, the underlying molecular mechanisms remain poorly understood. METHOD...BACKGROUND: Type 2 diabetes mellitus elevates the risk of developing tuberculosis, posing a substantial challenge to TB treatment and control. However, the underlying molecular mechanisms remain poorly understood. METHODS: Non-targeted metabolomics analysis of serum samples collected from 20 patients with concurrent DM and TB (DM-TB) and 20 patients with TB only (TB) was performed using UPLC-MS/MS. Logistic regressions were applied to evaluate the associations between the metabolites and lung pathology. RESULTS: Pathway analysis revealed that TCA cycle, steroidogenesis, ketone body metabolism were the key pathways enriched in the differential metabolites distinguishing DM-TB from TB patients. Multivariable logistic regression analysis suggested, high levels of C18 Sphingosine (OR = 2.207, 95% CI = 1.049-4.645), L-Saccharopine (OR = 4.407, 95% CI = 1.045-18.582) and arachidonic acid (AA) (OR = 2.845, 95% CI = 1.272-6.366) were associated with an increased risk of extensive lung lesions. In addition, C18 Sphingosine (OR = 3.596, 95% CI = 1.239-10.440) and AA levels (OR = 3.306, 95% CI = 1.200-9.112) were positively associated with the risk of prevalent multiple cavities. CONCLUSIONS: Our results suggests that lipid metabolism disorders are significantly associated with severe lung lesions in TB-DM patients, suggest a potential link to exacerbated inflammatory and immunomodulatory responses.
Tuberculosis (TB) frequently coexists with chronic conditions such as diabetes, HIV, malnutrition, sarcoidosis, and lung cancer. These comorbidities profoundly reshape the host immune, metabolic, and tissue environments,...Tuberculosis (TB) frequently coexists with chronic conditions such as diabetes, HIV, malnutrition, sarcoidosis, and lung cancer. These comorbidities profoundly reshape the host immune, metabolic, and tissue environments, disrupting pathways that govern Mycobacterium tuberculosis (Mtb) containment and altering responses to both antimicrobial and host-directed therapies. Yet, most TB drug discovery pipelines continue to rely on "healthy-host" experimental models, overlooking the biological heterogeneity of the populations most affected by the disease. In this review, we argue that TB should be reframed not as a single disease entity, but as a spectrum of comorbidity-modulated disease mechanisms shaped by host health status. We synthesize mechanistic evidence demonstrating how comorbidities rewire macrophage function, immune signaling, metabolic programs, autophagy, granuloma architecture, and drug distribution and accessibility, with direct consequences for pharmacokinetics, pharmacodynamics, and treatment efficacy. Using the framework of "One Drug, Multiple Disease Mechanisms," we highlight opportunities to repurpose drugs developed for comorbid conditions-such as metabolic, cardiovascular, neuropsychiatric, and oncologic agents-as cross-cutting host-directed interventions in TB. We further discuss emerging experimental and computational strategies, including comorbidity-mimicking cellular systems, organoids, CRISPR-based screens, dual-disease animal models, and network-level and systems-biology pathway modeling, to embed host complexity into TB target discovery and validation. By embracing host heterogeneity rather than filtering it out, TB research can prioritize resilient and translationally robust targets and advance therapeutic strategies that are effective across diverse, comorbidity-burdened patient populations.
Rv2660c is present in both Mycobacterium tuberculosis (M. tuberculosis) and Mycobacterium bovis (M. bovis) and has been reported as a protective antigen. In this study, Rv2660c was linked with another M. tuberculosis spe...Rv2660c is present in both Mycobacterium tuberculosis (M. tuberculosis) and Mycobacterium bovis (M. bovis) and has been reported as a protective antigen. In this study, Rv2660c was linked with another M. tuberculosis specific antigen Rv2645, which is present in M. tuberculosis but not in M. bovis, to construct a fusion protein Rv2645-Rv2660c (LT25 in short). LT25 failed to confer protection following M. bovis challenge. Structural predictions via AlphaFold 3 revealed that LT25 retained the native conformation of Rv2645, whereas Rv2660c underwent significant conformational alterations. LT25 elicited strong Rv2645 and Rv2660c-specific cellular immune responses and high levels of Rv2645-specific antibodies, but lower levels of Rv2660c-specific antibodies than Rv2660c did. Further investigation showed that the Rv2660c-induced polyclonal antibody possessed stronger phagocytosis-promoting activity than LT25-induced antibodies did. These findings suggest that the structural integrity of Rv2660c is essential for its protective function.
In pleural tuberculosis (TB), pleural effusion is typically characterized by a lymphocytic-rich exudate; however, neutrophils and macrophages predominate during the initial phase of infection. These immune cells are know...In pleural tuberculosis (TB), pleural effusion is typically characterized by a lymphocytic-rich exudate; however, neutrophils and macrophages predominate during the initial phase of infection. These immune cells are known to combat pathogens via phagocytosis, degranulation, and formation of extracellular traps (ETs)-a recently recognized defence mechanism. Although in-vitro and in-vivo studies have demonstrated ETs formation during Mycobacterium tuberculosis infection, their presence in clinical samples of patients with pleural TB has not been previously demonstrated. In this study, we employed multiple methods to demonstrate the presence of ETs in tuberculous pleural effusion (TPE) from patients with confirmed TB (Xpert MTB/RIF-positive/culture-positive/pleural biopsy Ziehl-Neelsen-stainingacid fast bacilli-positive). Immunofluorescence microscopy revealed DNA co-localized with ET markers like myeloperoxidase, neutrophil elastase, and citrullinated histones-confirming ETs in TPE samples, and this was further supported by immunoblotting. Proteomic analysis of TPE samples revealed the presence of key ET-associated proteins, including histones, myeloperoxidase, matrix metalloproteinase-9, and the S100A8/A9 complex. Protein-protein interaction and gene ontology analyses revealed that these proteins are involved in ET-related biological processes, such as neutrophil degranulation and collagen degradation. To the best of our knowledge, this is the first clinical evidence of ETs in TPE, suggesting a potential role in the immunopathogenesis of pleural TB.
Cyclic dinucleotides (CDNs), particularly cyclic di-AMP (c-di-AMP) and cyclic di-GMP (c-di-GMP), are now recognised as major second messengers that governs critical physiological and pathogenic processes in Mycobacterium...Cyclic dinucleotides (CDNs), particularly cyclic di-AMP (c-di-AMP) and cyclic di-GMP (c-di-GMP), are now recognised as major second messengers that governs critical physiological and pathogenic processes in Mycobacterium tuberculosis. Beyond their central roles in cell wall homeostasis, DNA repair, metabolism, stress responses, and virulence, these signaling molecules also strongly influence host immunity through activation of the cGAS-STING pathway, generating significant interest in their potential as vaccine adjuvants. This review synthesizes recent progress in understanding CDN biosynthesis and degradation, the identification of new effector and receptor proteins, and the expanding regulatory networks governed by these molecules in mycobacteria. Emerging findings highlight the essential role of c-di-AMP in bacterial growth and genome integrity surveillance, the established involvement of c-di-GMP in lifestyle transitions, and the potent immunomodulatory properties of both CDNs. Targeting CDN signaling pathways or harnessing their immune-stimulatory functions, offers promising avenues for developing next-generation antimycobacterial therapeutics and improved vaccine strategies. This review integrates current advances, highlights recent breakthroughs, and outlines future challenges in decoding CDN signaling in mycobacteria.
Signorelli F, Marè C, De Matteis G
… +13 more, Grandoni F, Orrù L, Lamontanara A, Schiavo L, Donniacuo A, Cerrone P, Spoleto C, De Carlo E, Galiero G, Iovane G, Paciello O, Martucciello A, Napolitano F
Bovine tuberculosis (bTB), caused by Mycobacterium bovis, remains a major zoonosis impacting livestock productivity and public health. To characterize species-specific immune responses and identify transcriptional biomar...Bovine tuberculosis (bTB), caused by Mycobacterium bovis, remains a major zoonosis impacting livestock productivity and public health. To characterize species-specific immune responses and identify transcriptional biomarkers of infection, we quantified the expression of 45 immune-related genes in peripheral blood from naturally infected cattle and Mediterranean buffalo. Differential and multivariate analyses revealed distinct species- and state-specific transcriptional signatures. Sixteen genes were significantly modulated in cattle, with Canonical Discriminant Analysis (CDA) defining a promising diagnostic signature marked by upregulation of IFNG, CASP8, CASP1, and CD83, and downregulation of IL10, CXCR2, and MMP9. In buffaloes, 22 genes were differently expressed across the three clinical groups. The first canonical function (Can1), driven positively by IFNG and IL12B and negatively by MMP9 and TLR5, effectively separated healthy from infected and affected animals, while the second function (Can2) distinguished the affected status through marked upregulation of CXCL1 and IL6. The species-specific immune signatures uncovered underscore the importance of developing tailored biomarker panels for improved diagnosis and surveillance of bTB in different livestock species.
Tuberculosis (TB) is still a serious global public health problem, affecting mostly vulnerable populations such as the population deprived of liberty (PDL). Brazil has the third-largest prison population in the world, an...Tuberculosis (TB) is still a serious global public health problem, affecting mostly vulnerable populations such as the population deprived of liberty (PDL). Brazil has the third-largest prison population in the world, and the state of São Paulo has the largest PDL in the country. This study investigated the molecular epidemiology of TB in individuals from the general population (GP) and the PDL belonging to two Epidemiological Surveillance Groups in the state of São Paulo. A total of 522 Mycobacterium tuberculosis isolates were analyzed using MIRU-VNTR genotyping (15 and 24-loci) and multiplex PCR for detection of RD/RD174 deletions. Clustering rates were 14.7% for 15-loci and 12.8% for 24-loci. The RD sublineage was identified in 26.8% of isolates, showing an association with alcohol use disorder (OR = 1.84; 95% CI 1.17-2.88) and illiteracy (OR = 3.40; 95% CI 1.09-10.58). Clusters with 100% similarity between GP and PDL isolates were observed, indicating intra- and extramural transmission within prison units. These findings highlight the importance of integrated surveillance and control strategies, with strengthened active case finding in both populations, in order to interrupt transmission chains and reduce the TB burden. In addition, further high-resolution genomic studies involving the PDL are recommended, focusing on complete genome sequencing integrated with epidemiological, spatial, and socio-environmental data, which will contribute to the improvement of TB surveillance and control strategies in highly vulnerable contexts.
Drug tolerance in Mycobacterium tuberculosis (Mtb) significantly undermines the success of antimycobacterial therapy. Redox active compounds such as Vitamin C (VitC) and iron can modulate drug efficacy, and in the presen...Drug tolerance in Mycobacterium tuberculosis (Mtb) significantly undermines the success of antimycobacterial therapy. Redox active compounds such as Vitamin C (VitC) and iron can modulate drug efficacy, and in the present study we evaluated their effect on Rifampicin (RIF) mediated killing of Mycobacterium tuberculosis H37Ra (Mtb-Ra). We also studied the cellular reactive oxygen species (ROS) levels and expression of stress-response and drug target genes, and host cell cytotoxicity. ROS measurements revealed that treating Mtb-Ra with RIF and VitC led to ROS levels mostly being elevated, with iron supplementation leading to further increase. Survival analysis showed that VitC increased the killing by RIF in a dose-dependent manner, with iron potentiating this effect. Cytotoxicity studies showed that high iron and its combinations with VitC and RIF showed comparable cytotoxicity. The qRT-PCR study demonstrated RIF induced up-regulation of recA, dnaE2 and rpoB, with VitC and iron further amplifying recA and dnaE2 expression. Conversely, lexA was down-regulated in the presence of VitC and more strongly with iron, indicating induction of bacterial SOS response. Interestingly, rpoB expression, while up-regulated with RIF and VitC, was suppressed by iron. These findings suggest that VitC and iron-induced disruption of Mtb homeostasis can enhance RIF efficacy.
Tuberculosis (TB) is a curable infectious disease that requires prolonged treatment with multiple antibiotics. To better understand how the immune system contributes to the clearance of Mycobacterium tuberculosis (Mtb),...Tuberculosis (TB) is a curable infectious disease that requires prolonged treatment with multiple antibiotics. To better understand how the immune system contributes to the clearance of Mycobacterium tuberculosis (Mtb), in vitro assays are essential for monitoring functional immune changes during infection, therapy, and following vaccination. In this study, we investigated whether mycobacterial growth control in thirty patients with TB disease changes over the course of treatment. Comprehensive immune profiling of peripheral blood mononuclear cells (PBMCs), using a 30-color spectral flow cytometry panel, identified dynamic shifts in immune cell subsets related to functional activity. Notably, in addition to memory and effector T cells, a subset of naive B cells changed during treatment. Most sera contained antibodies binding to purified protein derivative (PPD) and Mtb-specific antigens ESAT-6/CFP-10, and enhanced phagocytic activity. A functional mycobacterial growth inhibition assay (MGIA) revealed growth control, which appeared to be heterogeneous but generally sustained throughout longitudinal follow-up. We conclude that T and B cell responses change in response to antibiotic treatment of TB disease, but that mycobacterial growth control capacity is a property of the individual, which is not influenced by disease activity or antibiotic treatment.
Tuberculosis (Edinb)
· 2026 May · PMID 41926880
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Tuberculosis (TB) remains a major global health challenge due in part to limitations in rapid and affordable diagnostics. Current diagnostic methods are time-intensive and often inaccessible in resource-limited settings,...Tuberculosis (TB) remains a major global health challenge due in part to limitations in rapid and affordable diagnostics. Current diagnostic methods are time-intensive and often inaccessible in resource-limited settings, emphasizing the urgent need for rapid, low-cost screening approaches. One promising strategy involves the analysis of volatile molecules associated with TB-infection. In this study (n = 100) we identify 14 sputum-derived volatiles and utilize them to construct a machine learning model that classifies samples by TB status with a sensitivity of 90% and a specificity of 86% across cross-validation folds. The resulting profile provides a foundation for further biomarker validation studies with an expanded sample size and the development of non-invasive breath diagnostics.
BACKGROUND: Spinal tuberculosis (STB) is an infectious disease caused by Mtb with unclear diagnosis and molecular mechanisms. Autophagy is reported to be associated with the pathology of spinal tuberculosis. The present...BACKGROUND: Spinal tuberculosis (STB) is an infectious disease caused by Mtb with unclear diagnosis and molecular mechanisms. Autophagy is reported to be associated with the pathology of spinal tuberculosis. The present study intends to elucidate the role of autophagy-related miRNAs and genes in STB. METHODS: Core miRNAs were identified through WGCNA and differential analysis. A total of 113 machine learning algorithms were used to develop a diagnostic model. Target genes were predicted and overlapped by TargetScan, miRDB, and miRTarBase. The two-sample Mendelian randomization analysis was utilized to explore the association between genes and tuberculosis. RESULTS: Nine autophagy-related miRNAs were identified. The GBM model yielded the best performance with the highest AUC (0.816). A signature comprising eight miRNAs, specifically miR-27b-3p and miR-27a-3p, was constructed accordingly. A nomogram was established to facilitate clinical implementation. ZFHX3 gene was indicated to be significantly associated with sequelae tuberculosis. Notably, the ZFHX3/miR-27 axis has never been reported in the realm of tuberculosis. CONCLUSIONS: The present research established an optimal machine learning model to predict the possibility of STB, which might provide valuable insights into the diagnosis and treatment of STB. ZFHX3/miR-27 may serve as a novel potential molecular pathway in Mtb pathophysiology.