Inflammasome Activation Differences Underpin Different Mycobacterium tuberculosis Infection Outcomes

Abstract

The clinical outcome of Mycobacterium tuberculosis (Mtb) infection ranges from latent/nonprogressive disease to active/progressive tuberculosis (TB), but the cellular events contributing to these variable outcomes remain unknown. Here, we report that progressive Mtb infection is associated with upregulation of guanylate-binding protein-1 (GBP1), hypoxia-inducible factor-1 alpha (HIF-1α), and elevated NLR family pyrin domain-containing (NLRP3) inflammasome activation pathways. Using rabbit lungs and primary rabbit and human macrophages, as well as human monocytic THP-1-derived macrophages for infection with Mtb strains (H₃₇Rv, HN878, or CDC1551) that differ in virulence, we show that NLRP3 inflammasome activation by HIF-1α and GBP1 leads to elevated mitochondrial stress, apoptosis, and necrosis during progressive infection by HN878. These biological functions and pathways are dampened during nonprogressive TB in rabbit lungs, and in primary rabbit and human macrophages infected by CDC1551. These findings are consistent with and confirmed by Mtb infection studies of macrophages knocked down for HIF-1α or GBP1 expression. Our study indicates that differences in HIF-1α- and GBP1-mediated NLRP3 inflammasome activation influence the outcome of Mtb infection in the host.

Keywords

host-pathogen interactions / immune response to infection / tuberculosis

References

[1] World Health Organization. Global Tuberculosis Report. World Health Organization; 2023. Accessed July 4, 2024. https://www.who.int/teams/global-programme-on-tuberculosis-and-lung-health/tb-reports/global-tuberculosis-report-2024.

[2] H. Chae and S. J. Shin, “Importance of Differential Identification of Mycobacterium Tuberculosis Strains for Understanding Differences in Their Prevalence, Treatment Efficacy, and Vaccine Development,” Journal of Microbiology (Seoul, Korea) 56, no. 5 (2018): 300–311.

[3] I. Comas and S. Gagneux, “A Role for Systems Epidemiology in Tuberculosis Research,” Trends in Microbiology 19, no. 10 (2011): 492–500.

[4] G. Kallenius, M. Correia-Neves, H. Buteme, B. Hamasur, and S. B. Svenson, “Lipoarabinomannan, and Its Related Glycolipids, Induce Divergent and Opposing Immune Responses to Mycobacterium Tuberculosis Depending on Structural Diversity and Experimental Variations,” Tuberculosis (Edinburgh, Scotland) 96 (2016): 120–130.

[5] M. Smet, C. Pollard, A. De Beuckelaer, et al., “Mycobacterium Tuberculosis-Associated Synthetic Mycolates Differentially Exert Immune Stimulatory Adjuvant Activity,” European Journal of Immunology 46, no. 9 (2016): 2149–2154.

[6] C. E. Stamm, A. C. Collins, and M. U. Shiloh, “Sensing of Mycobacterium Tuberculosis and Consequences to both Host and bacillus,” Immunological Reviews 264, no. 1 (2015): 204–219.

[7] L. D. Tientcheu, A. Koch, M. Ndengane, G. Andoseh, B. Kampmann, and R. J. Wilkinson, “Immunological Consequences of Strain Variation Within the Mycobacterium Tuberculosis Complex,” European Journal of Immunology 47, no. 3 (2017): 432–445.

[8] Y. C. Manabe, A. K. Kesavan, J. Lopez-Molina, et al., “The Aerosol Rabbit Model of TB Latency, Reactivation and Immune Reconstitution Inflammatory Syndrome,” Tuberculosis (Edinburgh, Scotland) 88, no. 3 (2008): 187–196.

[9] C. Manca, L. Tsenova, A. Bergtold, et al., “Virulence of a Mycobacterium Tuberculosis Clinical Isolate in Mice Is Determined by Failure to Induce Th1 Type Immunity and Is Associated With Induction of IFN-alpha /Beta,” PNAS 98, no. 10 (2001): 5752–5757.

[10] S. E. Valway, M. P. Sanchez, T. F. Shinnick, et al., “An Outbreak Involving Extensive Transmission of a Virulent Strain of Mycobacterium Tuberculosis,” New England Journal of Medicine 338, no. 10 (1998): 633–639.

[11] M. Hanekom, N. C. Gey van Pittius, C. McEvoy, T. C. Victor, P. D. Van Helden, and R. M. Warren, “Mycobacterium Tuberculosis Beijing Genotype: A Template for Success,” Tuberculosis (Edinburgh, Scotland) 91, no. 6 (2011): 510–523.

[12] M. S. Koo, S. Subbian, and G. Kaplan, “Strain Specific Transcriptional Response in Mycobacterium Tuberculosis Infected Macrophages,” Cell Communication and Signaling 10, no. 1 (2012): 2.

[13] C. Manca, L. Tsenova, C. E. Barry, et al., “Mycobacterium Tuberculosis CDC1551 Induces a More Vigorous Host Response in Vivo and in Vitro, but Is Not More Virulent Than Other Clinical Isolates,” Journal of Immunology 162, no. 11 (1999): 6740–6746.

[14] M. Merker, C. Blin, S. Mona, et al., “Evolutionary History and Global Spread of the Mycobacterium Tuberculosis Beijing Lineage,” Nature Genetics 47, no. 3 (2015): 242–249.

[15] M. B. Reed, P. Domenech, C. Manca, et al., “A Glycolipid of Hypervirulent Tuberculosis Strains That Inhibits the Innate Immune Response,” Nature 431, no. 7004 (2004): 84–87.

[16] S. Subbian, L. Tsenova, G. Yang, et al., “Chronic Pulmonary Cavitary Tuberculosis in Rabbits: A Failed Host Immune Response,” Open Biology 1, no. 4 (2011): 110016.

[17] P. Elkington, M. E. Polak, M. T. Reichmann, and A. Leslie, “Understanding the Tuberculosis Granuloma: The Matrix Revolutions,” Trends in Molecular Medicine 28, no. 2 (2022): 143–154.

[18] S. S. Ranjeet Kumar, “Immune Correlates of Non-Necrotic and Necrotic Granulomas in Pulmonary Tuberculosis: A Pilot Study,” Journal of Respiration 1, no. 4 (2021): 248–259.

[19] M. J. Kim, H. C. Wainwright, M. Locketz, et al., “Caseation of human Tuberculosis Granulomas Correlates With Elevated Host Lipid Metabolism,” EMBO Molecular Medicine 2, no. 7 (2010): 258–274.

[20] J. C. Pena and W. Z. Ho, “Monkey Models of Tuberculosis: Lessons Learned,” Infection and Immunity 83, no. 3 (2015): 852–862.

[21] M. Silva Miranda, A. Breiman, S. Allain, F. Deknuydt, and F. Altare, “The Tuberculous Granuloma: An Unsuccessful Host Defence Mechanism Providing a Safety Shelter for the Bacteria?,” Clinical & Developmental Immunology 2012 (2012): 139127.

[22] S. Subbian, L. Tsenova, M. J. Kim, et al., “Lesion-Specific Immune Response in Granulomas of Patients With Pulmonary Tuberculosis: A Pilot Study,” PLoS ONE 10, no. 7 (2015): e0132249.

[23] S. Subbian, L. Tsenova, and P. O'Brien, “Spontaneous Latency in a Rabbit Model of Pulmonary Tuberculosis,” American Journal of Pathology 181, no. 5 (2012): 1711–1724.

[24] H. Z. Imtiyaz and M. C. Simon, “Hypoxia-Inducible Factors as Essential Regulators of Inflammation,” Current Topics in Microbiology and Immunology 345 (2010): 105–120.

[25] A. Palazon, A. W. Goldrath, V. Nizet, and R. S. Johnson, “HIF Transcription Factors, Inflammation, and Immunity,” Immunity 41, no. 4 (2014): 518–528.

[26] N. C. Denko, “Hypoxia, HIF1 and Glucose Metabolism in the Solid Tumour,” Nature Reviews Cancer 8, no. 9 (2008): 705–713.

[27] Q. Li, Y. Xie, Z. Cui, et al., “Activation of Hypoxia-inducible Factor 1 (Hif-1) Enhanced Bactericidal Effects of Macrophages to Mycobacterium Tuberculosis,” Tuberculosis (Edinburgh, Scotland) 126 (2021): 102044.

[28] M. Osada-Oka, N. Goda, H. Saiga, et al., “Metabolic Adaptation to Glycolysis Is a Basic Defense Mechanism of Macrophages for Mycobacterium Tuberculosis Infection,” International Immunology 31, no. 12 (2019): 781–793.

[29] J. J. Huang, J. Xia, L. L. Huang, and Y. C. Li, “HIF‑1alpha Promotes NLRP3 Inflammasome Activation in Bleomycin‑Induced Acute Lung Injury,” Molecular Medicine Reports 20, no. 4 (2019): 3424–3432.

[30] N. Challagundla and B. Saha, “Agrawal-Rajput R. Insights Into Inflammasome Regulation: Cellular, Molecular, and Pathogenic Control of Inflammasome Activation,” Immunologic Research 70, no. 5 (2022): 578–606.

[31] X. Liu, Z. Zhang, J. Ruan, et al., “Inflammasome-activated Gasdermin D Causes Pyroptosis by Forming Membrane Pores,” Nature 535, no. 7610 (2016): 153–158.

[32] K. Schroder and J. Tschopp, “The Inflammasomes,” Cell 140, no. 6 (2010): 821–832.

[33] D. Zheng, T. Liwinski, and E. Elinav, “Inflammasome Activation and Regulation: Toward a Better Understanding of Complex Mechanisms,” Cell Discovery 6 (2020): 36.

[34] B. M. Cumming, K. W. Addicott, J. H. Adamson, and A. J. Steyn, “Mycobacterium Tuberculosis Induces Decelerated Bioenergetic Metabolism in Human Macrophages,” Elife 7 (2018): e39169.

[35] K. L. Patrick and R. O. Watson, “Mitochondria: Powering the Innate Immune Response to Mycobacterium Tuberculosis Infection,” Infection and Immunity 89, no. 4 (2021): e00687-20.

[36] A. T. Honkala, D. Tailor, and S. V. Malhotra, “Guanylate-Binding Protein 1: An Emerging Target in Inflammation and Cancer,” Frontiers in Immunology 10 (2019): 3139.

[37] B. H. Kim, A. R. Shenoy, P. Kumar, R. Das, S. Tiwari, and J. D. MacMicking, “A family of IFN-Gamma-Inducible 65-kD GTPases Protects Against Bacterial Infection,” Science 332, no. 6030 (2011): 717–721.

[38] S. M. Man, D. E. Place, T. Kuriakose, and T. D. Kanneganti, “Interferon-Inducible Guanylate-Binding Proteins at the Interface of Cell-Autonomous Immunity and Inflammasome Activation,” Journal of Leukocyte Biology 101, no. 1 (2017): 143–150.

[39] D. Pilla-Moffett, M. F. Barber, G. A. Taylor, and J. Coers, “Interferon-Inducible GTPases in Host Resistance, Inflammation and Disease,” Journal of Molecular Biology 428, no. 17 (2016): 3495–3513.

[40] A. R. Shenoy, D. A. Wellington, P. Kumar, et al., “GBP5 promotes NLRP3 Inflammasome Assembly and Immunity in Mammals,” Science 336, no. 6080 (2012): 481–485.

[41] C. C. Ngo and S. M. Man, “Mechanisms and Functions of Guanylate-Binding Proteins and Related Interferon-Inducible GTPases: Roles in Intracellular Lysis of Pathogens,” Cellular Microbiology 19, no. 12 (2017).

[42] S. A. Nicholas, V. V. Bubnov, I. M. Yasinska, and V. V. Sumbayev, “Involvement of Xanthine Oxidase and Hypoxia-Inducible Factor 1 in Toll-Like Receptor 7/8-Mediated Activation of Caspase 1 and Interleukin-1beta,” Cellular and Molecular Life Sciences 68, no. 1 (2011): 151–158.

[43] A. Nisa, F. C. Kipper, D. Panigrahy, S. Tiwari, A. Kupz, and S. Subbian, “Different Modalities of Host Cell Death and Their Impact on Mycobacterium Tuberculosis Infection,” American Journal of Physiology Cell Physiology 323, no. 5 (2022): C1444–C1474.

[44] A. Diatlova, N. Linkova, A. Lavrova, et al., “Molecular Markers of Early Immune Response in Tuberculosis: Prospects of Application in Predictive Medicine,” International Journal of Molecular Sciences 24, no. 17 (2023): 13261.

[45] J. Ghanavi, P. Farnia, P. Farnia, and A. A. Velayati, “The Role of Interferon-Gamma and Interferon-Gamma Receptor in Tuberculosis and Nontuberculous Mycobacterial Infections,” International Journal of Mycobacteriology 10, no. 4 (2021): 349–357.

[46] A. J. Olive, C. M. Smith, C. E. Baer, J. Coers, and C. M. Sassetti, “Mycobacterium Tuberculosis Evasion of Guanylate Binding Protein-Mediated Host Defense in Mice Requires the ESX1 Secretion System,” International Journal of Molecular Sciences 24, no. 3 (2023): 2861.

[47] K. Tretina, E. S. Park, A. Maminska, and J. D. MacMicking, “Interferon-Induced Guanylate-Binding Proteins: Guardians of Host Defense in Health and Disease,” Journal of Experimental Medicine 216, no. 3 (2019): 482–500.

[48] Q. Jiang, X. Geng, J. Warren, et al., “Hypoxia Inducible Factor-1alpha (HIF-1alpha) Mediates NLRP3 Inflammasome-Dependent-Pyroptotic and Apoptotic Cell Death Following Ischemic Stroke,” Neuroscience 448 (2020): 126–139.

[49] E. Meunier, M. S. Dick, R. F. Dreier, et al., “Caspase-11 Activation Requires Lysis of Pathogen-containing Vacuoles by IFN-induced GTPases,” Nature 509, no. 7500 (2014): 366–370.

[50] X. Ouyang, A. Ghani, A. Malik, et al., “Adenosine Is Required for Sustained Inflammasome Activation via the A(2)A Receptor and the HIF-1alpha Pathway,” Nature Communications 4 (2013): 2909.

[51] D. M. Pilla, J. A. Hagar, A. K. Haldar, et al., “Guanylate Binding Proteins Promote Caspase-11-dependent Pyroptosis in Response to Cytoplasmic LPS,” Proceedings of National Academy of Sciences 111, no. 16 (2014): 6046–6051.

[52] A. M. Dannenberg and F. M. Collins, “Progressive Pulmonary Tuberculosis Is Not Due to Increasing Numbers of Viable bacilli in Rabbits, Mice and guinea Pigs, but Is Due to a Continuous Host Response to Mycobacterial Products,” Tuberculosis (Edinburgh, Scotland) 81, no. 3 (2001): 229–242.

[53] A. Dusthackeer, M. Balasubramanian, G. Shanmugam, et al., “Differential Culturability of Mycobacterium Tuberculosis in Culture-Negative Sputum of Patients with Pulmonary Tuberculosis and in a Simulated Model of Dormancy,” Frontiers in Microbiology 10 (2019): 2381.

[54] R. L. Hunter, “The Pathogenesis of Tuberculosis-The Koch Phenomenon Reinstated,” Pathogens 9, no. 10 (2020): 813.

[55] S. Verma, K. Bhatt, A. Lovey, et al., “Transmission Phenotype of Mycobacterium Tuberculosis Strains Is Mechanistically Linked to Induction of Distinct Pulmonary Pathology,” Plos Pathogens 15, no. 3 (2019): e1007613.

[56] Y. C. Manabe, A. M. Dannenberg, and S. K. Tyagi, “Different Strains of Mycobacterium Tuberculosis Cause Various Spectrums of Disease in the Rabbit Model of Tuberculosis,” Infection and Immunity 71, no. 10 (2003): 6004–6011.

[57] L. Tsenova, D. Fallows, A. Kolloli, et al., “Inoculum Size and Traits of the Infecting Clinical Strain Define the Protection Level Against Mycobacterium Tuberculosis Infection in a Rabbit Model,” European Journal of Immunology 50, no. 6 (2020): 858–872.

[58] M. S. Cardoso, T. M. Silva, M. Resende, R. Appelberg, and M. Borges, “Lack of the Transcription Factor Hypoxia-Inducible Factor 1alpha (HIF-1alpha) in Macrophages Accelerates the Necrosis of Mycobacterium Avium-Induced Granulomas,” Infection and Immunity 83, no. 9 (2015): 3534–3544.

[59] R. Kumar, A. Kolloli, S. Subbian, D. Kaushal, L. Shi, and S. Tyagi, “Imaging the Architecture of Granulomas Induced by Mycobacterium Tuberculosis Infection With Single-Molecule Fluorescence in Situ Hybridization,” Journal of Immunology 213, no. 4 (2024): 526–537.

[60] M. Resende, C. M. Ferreira, A. M. Barbosa, et al., “Myeloid HIF-1alpha Regulates Pulmonary Inflammation During Experimental Mycobacterium Tuberculosis Infection,” Immunology 159, no. 1 (2020): 121–129.

[61] M. M. Costa Franco, F. Marim, E. S. Guimaraes, et al., “Brucella Abortus Triggers a cGAS-Independent STING Pathway To Induce Host Protection That Involves Guanylate-Binding Proteins and Inflammasome Activation,” Journal of Immunology 200, no. 2 (2018): 607–622.

[62] W. Cui, E. Braun, W. Wang, et al., “Structural Basis for GTP-Induced Dimerization and Antiviral Function of Guanylate-binding Proteins,” Proceedings of National Academy of Sciences 118, no. 15 (2021): e2022269118.

[63] D. Fisch, H. Bando, B. Clough, et al., “Human GBP1 Is a Microbe-Specific Gatekeeper of Macrophage Apoptosis and Pyroptosis,” EMBO Journal 38, no. 13 (2019): e100926.

[64] M. T. R. Gomes, D. M. Cerqueira, E. S. Guimaraes, P. C. Campos, and S. C. Oliveira, “Guanylate-Binding Proteins at the Crossroad of Noncanonical Inflammasome Activation During Bacterial Infections,” Journal of Leukocyte Biology 106, no. 3 (2019): 553–562.

[65] B. H. Kim, J. D. Chee, C. J. Bradfield, E. S. Park, P. Kumar, and J. D. MacMicking, “Interferon-Induced Guanylate-Binding Proteins in Inflammasome Activation and Host Defense,” Nature Immunology 17, no. 5 (2016): 481–489.

[66] B. H. Kim, A. R. Shenoy, P. Kumar, C. J. Bradfield, and J. D. MacMicking, “IFN-inducible GTPases in Host Cell Defense,” Cell Host & Microbe 12, no. 4 (2012): 432–444.

[67] Q. Liu, D. Zhang, D. Hu, X. Zhou, and Y. Zhou, “The Role of Mitochondria in NLRP3 Inflammasome Activation,” Molecular Immunology 103 (2018): 115–124.

[68] F. V. Marinho, J. S. Fahel, A. de Araujo, et al., “Guanylate Binding Proteins Contained in the Murine Chromosome 3 Are Important to Control Mycobacterial Infection,” Journal of Leukocyte Biology 108, no. 4 (2020): 1279–1291.

[69] X. Qiu, H. Guo, J. Yang, Y. Ji, C. S. Wu, and X. Chen, “Down-regulation of Guanylate Binding Protein 1 Causes Mitochondrial Dysfunction and Cellular Senescence in Macrophages,” Scientific Reports 8, no. 1 (2018): 1679.

[70] J. C. Santos, D. Boucher, L. K. Schneider, et al., “Human GBP1 Binds LPS to Initiate Assembly of a Caspase-4 Activating Platform on Cytosolic Bacteria,” Nature Communications 11, no. 1 (2020): 3276.

[71] T. Shi, L. Huang, Y. Zhou, and J. Tian, “Role of GBP1 in Innate Immunity and Potential as a Tuberculosis Biomarker,” Scientific Reports 12, no. 1 (2022): 11097.

[72] S. Peer, H. S. Jabbal, P. Singh, M. PS, S. Kakkera, and P. Bhat, “A Case Report of Successful Percutaneous Aspiration, Injection, and Re-aspiration (PAIR) Technique for Treatment of Retrovesical Pelvic Hydatid Cyst,” Radiology Case Report 18, no. 1 (2023): 331–334.

[73] E. P. Amaral, S. Namasivayam, A. T. L. Queiroz, et al., “BACH1 promotes Tissue Necrosis and Mycobacterium Tuberculosis Susceptibility,” Nature Microbiology 9, no. 1 (2024): 120–135.

[74] F. J. Roca, L. J. Whitworth, S. Redmond, A. A. Jones, and L. Ramakrishnan, “TNF Induces Pathogenic Programmed Macrophage Necrosis in Tuberculosis Through a Mitochondrial-Lysosomal-Endoplasmic Reticulum Circuit,” Cell 178, no. 6 (2019): 1344–1361. e11.

[75] D. Bertheloot, E. Latz, and B. S. Franklin, “Necroptosis, Pyroptosis and Apoptosis: An Intricate Game of Cell Death,” Cellular and Molecular Immunology 18, no. 5 (2021): 1106–1121.

[76] A. Lam, R. Prabhu, C. M. Gross, L. A. Riesenberg, V. Singh, and S. Aggarwal, “Role of Apoptosis and Autophagy in Tuberculosis,” American Journal of Physiology Lung Cellular and Molecular Physiology 313, no. 2 (2017): L218–L229.

[77] S. M. Behar, M. Divangahi, and H. G. Remold, “Evasion of Innate Immunity by Mycobacterium Tuberculosis: Is Death an Exit Strategy?,” Nature Reviews Microbiology 8, no. 9 (2010): 668–674.

[78] R. Blomgran, L. Desvignes, V. Briken, and J. D. Ernst, “Mycobacterium Tuberculosis Inhibits Neutrophil Apoptosis, Leading to Delayed Activation of Naive CD4 T Cells,” Cell Host & Microbe 11, no. 1 (2012): 81–90.

[79] R. E. Butler, P. Brodin, J. Jang, et al., “The Balance of Apoptotic and Necrotic Cell Death in Mycobacterium Tuberculosis Infected Macrophages Is Not Dependent on Bacterial Virulence,” PLoS ONE 7, no. 10 (2012): e47573.

[80] C. Antimicrobial Resistance, “Global Burden of Bacterial Antimicrobial Resistance in 2019: A Systematic Analysis,” Lancet 399, no. 10325 (2022): 629–655.

[81] V. Cody, H. Shen, M. Shlyankevich, R. E. Tigelaar, J. L. Brandsma, and D. J. Hanlon, “Generation of Dendritic Cells From Rabbit Bone Marrow Mononuclear Cell Cultures Supplemented With hGM-CSF and hIL-4,” Veterinary Immunology and Immunopathology 103, no. 3-4 (2005): 163–172.

[82] M. Kumar, S. K. Sahu, R. Kumar, et al., “MicroRNA Let-7 Modulates the Immune Response to Mycobacterium Tuberculosis Infection via Control of A20, an Inhibitor of the NF-kappaB Pathway,” Cell Host & Microbe 17, no. 3 (2015): 345–356.

[83] R. Kumar, S. K. Sahu, M. Kumar, et al., “MicroRNA 17-5p Regulates Autophagy in Mycobacterium Tuberculosis-Infected Macrophages by Targeting Mcl-1 and STAT3,” Cellular Microbiology 18, no. 5 (2016): 679–691.

RIGHTS & PERMISSIONS

2025 The Author(s). MedComm published by Sichuan International Medical Exchange & Promotion Association (SCIMEA) and John Wiley & Sons Australia, Ltd.