OBJECTIVE: This study explores the link between immune cells and tuberculosis (TB) pathogenesis and progression, proposing diagnostic strategies based on immune microenvironment changes. METHODS: The CIBERSORT algorithm...OBJECTIVE: This study explores the link between immune cells and tuberculosis (TB) pathogenesis and progression, proposing diagnostic strategies based on immune microenvironment changes. METHODS: The CIBERSORT algorithm assessed immune cell infiltration in TB tissues, validated by routine blood tests. Differential expression analysis and WGCNA identified key genes and modules. GO and KEGG analyses elucidated biological functions. Machine learning pinpointed diagnostic biomarkers and built a predictive model. Further validation included GSVA, single-cell data, Mendelian randomization, and RT-qPCR. RESULTS: Analysis of the immune microenvironment in TB patients and healthy controls revealed monocytes as the predominant immune cell type. A total of 90 overlapping genes were identified through differential expression analysis and WGCNA. A diagnostic model incorporating MSRB2, CLEC4D, and ASGR2 was constructed using three distinct machine learning algorithms and logistic regression. Single-cell data analysis demonstrated that these three genes were predominantly expressed in mononuclear cells of TB patients. MR analysis further established a causal relationship between CLEC4D and an elevated risk of TB. CONCLUSION: We established a monocyte-based diagnostic model demonstrating robust predictive accuracy. MSRB2, CLEC4D, and ASGR2 represent promising therapeutic targets for TB immunotherapy, providing potential breakthroughs in diagnostic precision and treatment efficacy.
Tuberculosis is a serious public health concern on a global scale, which emphasises the critical need for quick and precise diagnostic and treatment response monitoring techniques. In this study, Luminex multiplex immuno...Tuberculosis is a serious public health concern on a global scale, which emphasises the critical need for quick and precise diagnostic and treatment response monitoring techniques. In this study, Luminex multiplex immunoassay was used to detect the concentrations of 37 host biomarkers in saliva samples from 46 patients newly diagnosed with active pulmonary tuberculosis (PTB) and 46 patients with other respiratory diseases (ORD). Multiple logistic regression and the area under the receiver operator characteristics curve (AUC) were used to evaluate the diagnostic accuracy of biomarkers, which showed significant differences between the 2 groups. This study reported that Fractalkine exhibited the highest diagnostic accuracy and excellent discriminatory power, with statistically significant results (p ≤ 0.05), an AUC of 0.91, 89.1 % sensitivity and 76.1 % specificity, highlighting its strong potential to distinguish PTB cases from ORD cases. Additionally, our study found that the median levels of IL-17A, IL-23, and VEGF were statistically significant (p ≤ 0.05). General discriminant analysis further identified Fractalkine, VEGF, GM-CSF, IL-23, and IL-1α as the top five most effective biomarkers for combinations. The backward elimination approach demonstrated the potential usefulness of a four-marker combination (Fractalkine + GM-CSF + IL-23 + IL-1α) as a confirmatory diagnostic tool by achieving the greatest overall diagnostic accuracy with an AUC of 0.94 and 91.3 % specificity. Thus, combining multiple markers with high discriminating power may improve diagnostic performance and subsequently provide a more accurate, non-invasive saliva-based PTB diagnostic tool.
Tuberculosis (TB) remains a global health challenge, necessitating deeper insights into the dynamics of extracellular bacterial populations within infected hosts. This study presents an in-host mathematical model that in...Tuberculosis (TB) remains a global health challenge, necessitating deeper insights into the dynamics of extracellular bacterial populations within infected hosts. This study presents an in-host mathematical model that incorporates hereditary and antimicrobial factors influencing TB progression. The biological feasibility of the model is established by analyzing the boundedness of solutions within a realistic parameter space. The equilibrium states, including the disease-free and endemic equilibria, are examined, revealing conditions under which the system remains locally asymptotically stable. Sensitivity analysis is conducted to determine the key parameters driving infection dynamics, providing insights into potential control strategies. Notably, the model exhibits a backward bifurcation, indicating the possibility of multiple stable states and suggesting that reducing the basic reproduction number R below unity may not be sufficient for disease eradication. These findings highlight the importance of targeted interventions to effectively control extracellular bacterial populations and mitigate TB infection.
Mycobacterium tuberculosis (Mtb) and severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) may induce immunopathology with extensive lung damage in hosts. To elucidate the dynamics of co-infection Mtb and SARS-CoV...Mycobacterium tuberculosis (Mtb) and severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) may induce immunopathology with extensive lung damage in hosts. To elucidate the dynamics of co-infection Mtb and SARS-CoV-2 and its impact on inflammatory mediators' expression, we conducted a study to evaluate A549, lung epithelial cells, as a potential model for hosting both pathogens simultaneously. Cell infection initiated with Mtb H37Rv and following a 24-h incubation period, the cells were then infected with SARS-CoV-2. After a 72 h incubation period, a precision test was conducted for both pathogens, and total RNA was extracted for subsequent analysis of gene expression by RT-qPCR of the target genes: IFN-γ, TNF-α, IL-6, and IL-1β. Additionally, the levels of IL-1β, IL-2, IL-4, IL-6, IL-10, IFN-γ, and TNF-α in the culture supernatants were measured. A549 cells are a stable and reliable cellular model for co-infection between Mycobacterium tuberculosis and SARS-CoV-2. Co-infection with both pathogens led to downregulation of IFN-γ, TNF-α, and IL-10, and upregulation of IL-6 and IL-1β compared to uninfected cells. A549 cells function as a cellular model for co-infection and seems a good model for elucidating host inflammatory responses in the initial site of infection.
BACKGROUND: Macrophages play central roles in the immunity response to infection of intracellular bacteria, including Mycobacterium tuberculosis (M.tb) in tuberculosis (TB). Methyltransferase-like 3 (METTL3) has been imp...BACKGROUND: Macrophages play central roles in the immunity response to infection of intracellular bacteria, including Mycobacterium tuberculosis (M.tb) in tuberculosis (TB). Methyltransferase-like 3 (METTL3) has been implicated in the macrophage regulation in TB, and this study intended to investigate the molecular mechanism of METTL3 with interferon regulatory factor-8 (IRF8) in TB using in vitro model established by M.tb-infected THP-1 macrophages. METHODS: RT-qPCR and Western blot were utilized to analyze mRNA and protein expression, respectively. Cell viability, proliferation, and apoptosis were examined through cell counting kit-8 assay, EdU assay, and flow cytometry/TUNEL assay. Inflammatory cytokines were detected via enzyme-linked immunosorbent assay. Methylated RNA Immunoprecipitation (MeRIP), RIP and Co-IP were performed to assess the interaction between genes. RESULTS: IRF8 knockdown alleviated injury and inflammation in M.tb-infected THP-1 macrophages. METTL3 enhanced IRF8 mRNA stability by inducing mA methylation. IGF2BP1 functioned as an mA reader to affect mA methylation of IRF8. The function of METTL3 in M.tb-induced THP-1 macrophages was attributed to the positive regulation of IRF8. IRF8 bound to TLR4 and METTL3 could regulate TLR4 expression via targeting IRF8. IRF8/TLR4 axis promoted M.tb-induced THP-1 cell injury and inflammation. TLR4/NF-kB pathway was activated by METTL3-mediated IRF8. CONCLUSION: These findings revealed that METTL3 expedited cell injury and inflammatory reaction in M.tb-infected THP-1 macrophages by inducing mA methylation of IRF8 to activate TLR4/NF-kB pathway.
BACKGROUND: The Gene Xpert MTB/RIF Ultra is a Cartridge-Based Nucleic Acid Amplification Test (CBNAAT) for rapid detection of Mycobacterium tuberculosis (MTB) and rifampicin (RIF) drug susceptibility testing (DST) patter...BACKGROUND: The Gene Xpert MTB/RIF Ultra is a Cartridge-Based Nucleic Acid Amplification Test (CBNAAT) for rapid detection of Mycobacterium tuberculosis (MTB) and rifampicin (RIF) drug susceptibility testing (DST) pattern. OBJECTIVE: 1)To compare the culture results of samples detected as a Trace on Gene Xpert MTB/RIF Ultra with MTB growth in liquid culture. 2) To compare the rifampicin indeterminate results from Gene Xpert MTB/RIF Ultra with MGIT RIF DST pattern. MATERIALS AND METHODS: A retrospective case record study was conducted from January 1, 2020 to August 31, 2024. Samples were evaluated using Gene Xpert MTB/RIF Ultra and MGIT liquid culture. MTB isolates from MGIT liquid culture were subjected to MGIT RIF DST. RESULT: A total of 1821 samples detected as trace in Gene Xpert MTB/RIF Ultra, 538 (29.54 %) grew in MGIT liquid culture, while 1283 (70.46 %) did not grow. Among the 538 Trace detected samples with RIF indeterminate results, a comparison with MGIT RIF DST revealed that 451 (83.83 %) were rifampicin sensitive, while 87 (16.17 %) were rifampicin resistance. 49 (9.10 %) of these were pulmonary samples, while 489 (90.90 %) were extrapulmonary samples. CONCLUSION: Gene Xpert MTB/RIF Ultra is more rapid, more sensitive, less specific and more cost-effective compared to MGIT liquid culture.
The management of Mycobacterium abscessus (MAB) infection presents formidable challenges, necessitating the development of innovative antimicrobial agents and more efficacious combination therapies. The objective of this...The management of Mycobacterium abscessus (MAB) infection presents formidable challenges, necessitating the development of innovative antimicrobial agents and more efficacious combination therapies. The objective of this study was to investigate the in vitro, intracellular, and in vivo antibacterial activity of sudapyridine (WX-081) in combination with clarithromycin (CLR), clofazimine (CFZ), rifabutin (RFB), and linezolid (LZD) against MAB. In vitro, the interaction between WX-081 and other drugs was assessed using the checkerboard method against MAB. Synergistic or additive effects were observed when combining WX-081 with other drugs against all strains tested. The intracellular activity of WX-081 in combination with CLR, CFZ, RFB, and LZD against the MAB reference strain in J774A.1 cells was evaluated. The combination of WX-081 and CFZ or RFB exhibited potent inhibitory activity against MAB in macrophages. To evaluate pharmacodynamics, immunosuppressed BALB/c mice were infected with MAB to establish a murine model for assessing two-drug and three-drug combinations. In vivo studies demonstrated that the combination of WX-081 with CLR or both CLR and CFZ displayed antibacterial activity and significantly prevented mouse mortality. Our study demonstrates that the concurrent use of WX-081 with CLR or CLR plus CFZ holds significant promise as a clinical intervention for MAB infection.
OBJECTIVES: NTB-3119M, a novel benzothiopyranone derivative identified through comprehensive drug development studies, was selected as a promising antituberculosis (anti-TB) candidate. This study systematically evaluated...OBJECTIVES: NTB-3119M, a novel benzothiopyranone derivative identified through comprehensive drug development studies, was selected as a promising antituberculosis (anti-TB) candidate. This study systematically evaluated its anti-TB efficacy in vitro and in vivo. METHODS: In vitro analyses encompassed antimicrobial susceptibility testing to determine minimum inhibitory concentrations (MICs) against Mycobacterium tuberculosis H37Rv, 10 drug-susceptible clinical isolates, and 30 multidrug-resistant (MDR) strains, alongside evaluations of minimal bactericidal concentrations (MBCs) using H37Rv and seven clinical isolates. Additionally, intracellular anti-mycobacterial activity was assessed in H37Rv-infected macrophages, and cytotoxicity was profiled through MTT assays on Vero cells. In vivo studies utilized acute and chronic murine tuberculosis infection models to investigate the dose-dependent efficacy of NTB-3119M (50 and 100 mg/kg) against H37Rv, with concurrent comparative histopathological analysis of lung and spleen tissues. RESULTS: NTB-3119M demonstrated superior in vitro potency against both drug-sensitive and drug-resistant M. tuberculosis strains compared to first-line agents, Isoniazid (INH), Rifampicin (RIF), Moxifloxacin (MOFX), Levofloxacin (LVFX), and Streptomycin (SM), exhibiting comparable activity to PBTZ169. Time-kill curves for NTB-3119M indicate its potent bactericidal activity. Meanwhile, No cytotoxicity was observed on Vero cells. Spontaneous resistant mutants of NTB-3119M appears at a frequency of 6.44 × 10 to 3.65 × 10. Most importantly, NTB-3119M demonstrates comparable activity of PBTZ169 and better bactericidal activity against M. tuberculosis than INH and RIF in the 50- and 100- mg/kg groups in acute and chronic murine models. CONCLUSION: Our research provided comprehensive evidence that NTB-3119M with increased water solubility and bioavailability based on previous research performed excellent anti-tuberculosis activity and less cytotoxicity, which effectively tackled the undesirable drug properties associated with previous benzothiopyrone derivatives. It is warranted that NTB-3119M, as a highly promising candidate anti tuberculosis drug, deserves further research and clinical trial.
Bovine tuberculosis susceptibility and pathogenesis are influenced by host immunity, which may be modulated by the host microbiota. While intestinal microbiota composition affects pulmonary diseases in humans, its role i...Bovine tuberculosis susceptibility and pathogenesis are influenced by host immunity, which may be modulated by the host microbiota. While intestinal microbiota composition affects pulmonary diseases in humans, its role in bovine tuberculosis remains unclear. This study explores the intestinal microbiota of cattle and its association with tuberculosis to better understand disease pathophysiology. A case-control study was conducted using small intestine content samples from cattle with and without tuberculosis, slaughtered in Rio Grande do Sul, Brazil. DNA extraction, 16S rRNA (V4) sequencing, and bioinformatics analyses assessed alpha and beta diversity, taxa characterization, differential abundance, and metabolic pathways. No significant differences in alpha and beta diversities between the groups were detected. However, the Bacillota/Bacteroidota ratio suggested dysbiosis associated with bovine tuberculosis. Differential abundance analysis showed that microorganisms belonging to the Bacillota phylum, the Eubacterium cellulosolvens group, Colidextribacter and Coprococcus genera were enriched in healthy cattle. Conversely, animals with tuberculosis showed higher abundances of Verrucomicrobiota phylum, Sphingomonadaceae and Eubacteriaceae families, and Solobacterium and Clostridia-UCG-014 genera. Moreover, metabolic pathways related to carbohydrate degradation were enriched in healthy animals, and biosynthetic pathways related to disease were enriched in tuberculosis animals. This study highlights associations between intestinal microbiota and bovine tuberculosis, providing insights into disease mechanisms.
BACKGROUND: Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), remains a primary global health concern, with significant long-term sequelae. Central nervous system TB (CNS-TB) is a clinical spectrum with enti...BACKGROUND: Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), remains a primary global health concern, with significant long-term sequelae. Central nervous system TB (CNS-TB) is a clinical spectrum with entities such as tuberculous meningitis and tuberculomas. Emerging evidence suggests that Mtb may directly or indirectly affect the hippocampus, a critical memory, learning, and cognition structure. OBJECTIVES: This review aims to summarize the current biological understanding of Mtb's impact on the hippocampus, elucidate its potential role in neurodegeneration, and introduce the concept of "Hippocampal syndrome secondary to tuberculosis (HSST)" as a novel chronic entity within the CNS-TB spectrum. METHODS: A comprehensive literature review was conducted to analyze how Mtb gains access to the brain, its neurotropism, and the resulting neuroinflammatory and neurodegenerative effects on the hippocampus. Data from clinical, histopathological, and experimental studies were evaluated to assess potential links between TB and cognitive impairment. RESULTS: Mtb can access the CNS through hematogenous dissemination, the "Trojan Horse" mechanism, or via the olfactory pathway, bypassing the blood-brain barrier (BBB). Once in the brain, Mtb induces chronic neuroinflammation and disrupts hippocampal structure. Studies suggest that TB increases the risk of Alzheimer's and Parkinson's diseases, with evidence of Mtb-driven amyloid-beta accumulation and neuronal loss. Furthermore, specific Mtb strains exhibit neurotropism and produce virulence factors that facilitate CNS invasion. CONCLUSIONS: Understanding the interaction between TB and neurocognitive disorders is critical for improving post-TB care. Recognizing HSST as a chronic condition within the CNS-TB spectrum may support early diagnosis and targeted interventions to mitigate long-term neurological consequences.
OBJECTIVES: Circulating cell-free DNA (cfDNA), including mitochondrial cfDNA (mt-cfDNA) and nuclear cfDNA (nu-cfDNA), are potential biomarkers for infectious diseases. However, cfDNA variations in TB contacts with latent...OBJECTIVES: Circulating cell-free DNA (cfDNA), including mitochondrial cfDNA (mt-cfDNA) and nuclear cfDNA (nu-cfDNA), are potential biomarkers for infectious diseases. However, cfDNA variations in TB contacts with latent tuberculosis infection (LTBI) and their potential link to a predominance of M1 monocyte polarization in LTBI remain unexplored. METHODS: Contacts of TB patients were screened for LTBI using the Interferon gamma (IFN-γ) release assay. Blood cfDNA was extracted, and mt-cfDNA and nu-cfDNA copy numbers were quantified by qPCR. cfDNA levels in the supernatant of M1-polarized THP-1-derived macrophages were measured. RESULTS: Levels of mt-cfDNA and nu-cfDNA were lower in the LTBI group (n = 76) than in the uninfected group (n = 58) (p = 0.012, and p < 0.001). The results were consistent in an age- and sex-matched analysis (n = 41 pairs). mt-cfDNA and nu-cfDNA levels were negatively associated with the TB-specific IFN-γ response (p = 0.009, p < 0.001). In the LTBI group, mt-cfDNA was negatively associated with the index case's bacterial burden (p = 0.045). In cell model, mt-cfDNA and nu-cfDNA levels in the supernatant from M1-polarized macrophage were lower than those from M2-polarized cells (p = 0.030, p = 0.045). CONCLUSIONS: TB contacts with LTBI had lower cfDNA levels, which correlated with index case infectivity. Reduced cfDNA in M1-polarized macrophages warrants further investigation into the mechanisms of cfDNA reduction in LTBI.
Wang X, Harper K, Sinha P
… +2 more, Johnson WE, Patil P
Tuberculosis (Edinb)
· 2025 Jul · PMID 40359654
·
Full text
BACKGROUND: Tuberculosis (TB) is the leading cause of infectious disease mortality worldwide. Numerous host blood-based gene expression signatures have been proposed in the literature as alternative tools for diagnosing...BACKGROUND: Tuberculosis (TB) is the leading cause of infectious disease mortality worldwide. Numerous host blood-based gene expression signatures have been proposed in the literature as alternative tools for diagnosing TB infection. However, the generalizability of these signatures to different patient contexts is not well-characterized. There is a pressing need for a well-curated database of TB gene expression studies for the systematic assessment of existing and newly developed TB gene signatures. RESULTS: We built curatedTBData, a manually-curated database of 49 human TB transcriptomic studies. This data resource is freely available through GitHub and as an R Bioconductor package that allows users to validate new and existing biomarkers without the challenges of harmonizing heterogeneous studies. We demonstrate the use of this data resource with cross-study comparisons for 72 human host blood-based TB gene signatures. For the comparison of subjects with active TB from healthy controls, 19 gene signatures had weighted mean AUC of 0.90 or greater, with the highest result of 0.94. In active TB disease versus latent TB infection, 7 gene signatures had weighted mean AUC of 0.90 or greater, with a maximum of 0.93. CONCLUSIONS: The curatedTBData data package offers a comprehensive resource of curated human blood-based gene expression and clinically annotated data. This resource will facilitate the development of new signatures that are generalizable across cohorts or more applicable to specific subsets of patients.
Mitochondria form an integral, yet frequently underappreciated, part of the immune response to Mycobacterium tuberculosis (M.tb), particularly within macrophages. Despite growing recognition for their role in infection a...Mitochondria form an integral, yet frequently underappreciated, part of the immune response to Mycobacterium tuberculosis (M.tb), particularly within macrophages. Despite growing recognition for their role in infection and immunity, studies investigating how mitochondrial DNA (mtDNA) variation influences host susceptibility to tuberculosis (TB) are limited. Notably, there are no studies in African-based populations, although Africans possess unparalleled human genetic diversity, including the earliest diverged mitochondrial haplogroups, and a high TB burden. This underrepresentation limits the discovery of novel ancestry-specific genetic loci associated with TB. In this review article, we describe the unique characteristics of mtDNA, highlight key mitochondrial functions relevant to macrophage responses during M.tb infection, and summarise published studies that investigate the role of host mtDNA variation in TB susceptibility. We further advocate for the inclusion of African populations in future studies to identify novel TB susceptibility genetic risk loci and expand the current knowledgebase on host TB susceptibility.
Accurate diagnostics are urgently needed for bovine TB - an economically devastating disease posing a re-emerging threat to veterinary and public health worldwide. MicroRNAs, post-transcriptional gene regulators involved...Accurate diagnostics are urgently needed for bovine TB - an economically devastating disease posing a re-emerging threat to veterinary and public health worldwide. MicroRNAs, post-transcriptional gene regulators involved in a range of biological processes and immunological pathways, have emerged as promising diagnostic biomarkers for numerous diseases. In human TB, microRNAs have been widely studied, but not much is currently known about their role in bovine TB. This study aimed to investigate the diagnostic potential of microRNAs in bTB through comprehensive analysis of their expression profiles in disparate states of M. bovis exposure. We used RNA-sequencing to characterize the global microRNAome of peripheral blood mononuclear cells from cattle that were either unvaccinated, BCG-vaccinated, unprotected or protected. A total of 468 microRNAs were detected across all samples, none of which were uniquely expressed in any group. Significant differential expression was observed for bta-miR-6122-3p, bta-miR-3533 and bta-miR29b in various comparisons. Subsequent target analysis of bta-miR-29b, a candidate biomarker in human tuberculosis, revealed that several genes (ACVR2A, PIK3R1, TBX21, TGFBR1 and TGFBR2) involved in a number of relevant T-cell and immune signaling pathways, were amongst the predicted targets. Overall, this study provides evidence that microRNAs could be promising novel biomarkers for bovine TB diagnostics.
Divens AM, Ryan KJ, Sette A
… +2 more, Lindestam Arlehamn CS, Robinson CM
Tuberculosis (Edinb)
· 2025 Jul · PMID 40328205
·
Full text
Tuberculosis (TB) is the leading cause of death due to a pathogen. The live-attenuated BCG vaccine is the only approved vaccine to prevent TB, but it fails to confer long-term protection. We hypothesize that the immunosu...Tuberculosis (TB) is the leading cause of death due to a pathogen. The live-attenuated BCG vaccine is the only approved vaccine to prevent TB, but it fails to confer long-term protection. We hypothesize that the immunosuppressive cytokine IL-27 may contribute to the inefficacies of the BCG vaccine. IL-27 is elevated in neonates, the population most commonly administered BCG, and levels increase further upon vaccination. IL-27 interferes with the phagolysosomal pathway, suggesting it may limit the diversity of antigens processed and presented to T cells. We hypothesized that in the absence of IL-27 signaling, BCG vaccination induces antigen-specific T cells that recognize a greater number of antigens and provide enhanced protection during M. tuberculosis (Mtb) challenge. CD3 T cells isolated from IL-27Rα KO mice vaccinated with BCG as neonates were more responsive to BCG and a Mtb peptide pool than T cells from vaccinated WT mice. Adoptive transfer of IL-27Rα KO T cells provided more consistent protection against Mtb than WT, but this was not observed in TCRα mice. A principal component analysis suggested a more consistent multifunctional cytokine response was associated IL-27Rα KO T cells. These findings enhance our understanding of IL-27 during neonatal vaccination and development of protective immunity.
Tuberculosis (TB) remains a global health challenge as annual mortality rate due to drug resistant TB is increasing exponentially. This is mostly associated with the delayed diagnosis of Multidrug-resistant (MDR) or late...Tuberculosis (TB) remains a global health challenge as annual mortality rate due to drug resistant TB is increasing exponentially. This is mostly associated with the delayed diagnosis of Multidrug-resistant (MDR) or latent TB. Effective management of TB demands development of novel immunological strategies, such as peptide-based/subunit vaccines that can stimulate specific immune responses. In this context, we evaluated the immunogenic potential of two Major Histocompatibility Complex (MHC) Class I/II-restricted peptides from Mycobacterium tuberculosis (M. tuberculosis): Rv2588c and Rv0148. The peptides were tested on T and monocyte populations from healthy donors and pulmonary TB (PTB) patients. Flow cytometry analysis revealed significant T cell activation in peripheral blood mononuclear cells (PBMC) from both groups. Enzyme-linked immunosorbent assay (ELISA) demonstrated a strong IFN-γ response, confirming effective T cell activation. Additionally, these peptides induced increased nitric oxide (NO) production in macrophages, indicating their role in activating the innate immune system. Overall, Rv2588c and Rv0148 peptides exhibited robust immunogenicity, stimulating both adaptive and innate immune responses in PBMCs from healthy and PTB individuals. These findings highlight their potential as promising TB vaccine candidates, paving the way for improved TB treatment and prevention strategies.
BACKGROUND: Ibuprofen (IBU) is a non-prescription analgesic drug from the non-steroidal anti-inflammatory drug class. It is widely used for treating pain, fever, and inflammation. Both the in silico and in vitro experime...BACKGROUND: Ibuprofen (IBU) is a non-prescription analgesic drug from the non-steroidal anti-inflammatory drug class. It is widely used for treating pain, fever, and inflammation. Both the in silico and in vitro experiments were performed to determine the antibacterial potentials of the IBU against Mycobacterium tuberculosis (Mtb). METHODS: The STITCH v.5 pipeline was used to analyze the interaction of IBU with the proteome of the Mtb H37Ra and H37Rv strains. The GFP-tagged Bacillus Calmette Guerin (BCG) and td-tomato-tagged Mtb H37Ra were used to determine the bacteriostatic and bactericidal activities of IBU. The IBU-treated THP-1-derived macrophages were infected by td-tomato-tagged Mtb H37Ra and wild-type BCG to analyze the effects of IBU on bacterial phagocytosis and apoptosis, respectively. RESULTS: The in-silico study revealed that the IBU interacts with Mtb proteins primarily involved in cellular process, metabolism, and virulence, and targets four virulent proteins of Mtb, e.g., Cyp-123, Cyp-126, Cyp-130, and Cyp-139 in the cytochrome p450 system. The increasing concentrations of IBU showed significant bacteriostatic activity against Mtb H37Ra in vitro, where the 100 μg/ml and 200 μg/ml concentrations especially led to almost complete bacterial growth arrest. The IBU treatment does not affect BCG-induced apoptosis of THP-1-derived macrophages, but significantly enhances bacterial uptake, especially at 100 μg/ml and 200 μg/ml concentrations. CONCLUSIONS: The IBU enhances Mtb uptake by macrophages and exhibits direct bacteriostatic activity in vitro.