Pyrazinamide susceptibility testing in &lt;I&gt;Mycobacterium tuberculosis&lt;/I&gt; using the fast resazurin microtiter assay plate

Pina, Rosângela Zampieri; Caleffi-Ferracioli, Katiany Rizzieri; Campanerut‐Sá, Paula Aline Zanetti; Ghiraldi-Lopez, L D; Pavan, Fernando Rogério; Siqueira, Vera Ld; Scodro, Regiane Bertin de Lima; Cardoso, Rosilene Fressatti

doi:10.5588/ijtld.16.0304

Cited by 3 publications

(2 citation statements)

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“…Unstimulated, iH37Rv-Mtb-and LPS-Stimulated hMDMs TB antimicrobials optimally elicit direct bactericidal effects on mycobacteria, both in cellular and axenic models of TB infection [5,[22][23][24][25][26][27]. Accordingly, examining how these antimicrobials affect immunometabolic profiles and mitochondrial function in parallel could offer some insight into how these antimicrobials potentially affect immune function [7], particularly in patients with active TB who are subjected to long-term antimicrobial treatment regimens.…”

Section: Tb Antimicrobials Differentially Affect Glycolysis and Oxidative Phosphorylation Levels Inmentioning

confidence: 99%

“…Use of the iH37Rv-Mtb strain was essential to negate any bactericidal effect of the TB antimicrobials on live Mtb. Concentrations of these TB antimicrobials, well documented in killing Mtb, were used as previously described [5,[22][23][24][25][26][27]. At 24 h post stimulation, levels of glycolysis and OXPHOS were determined, measured by assessing extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) respectively, utilising the Seahorse Extracellular Flux Analyser (Figure 1).…”

Section: Tb Antimicrobials Differentially Affect Glycolysis and Oxidative Phosphorylation Levels Inmentioning

confidence: 99%

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The Effect of Tuberculosis Antimicrobials on the Immunometabolic Profiles of Primary Human Macrophages Stimulated with Mycobacterium tuberculosis

Cahill

Cox

O’Connell

et al. 2021

IJMS

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Tuberculosis (TB) remains a global health challenge. Patients with drug-sensitive and drug-resistant TB undergo long, arduous, and complex treatment regimens, often involving multiple antimicrobials. While these drugs were initially implemented based on their bactericidal effects, some studies show that TB antimicrobials can also directly affect cells of the immune system, altering their immune function. As use of these antimicrobials has been the mainstay of TB therapy for over fifty years now, it is more important than ever to understand how these antimicrobials affect key pathways of the immune system. One such central pathway, which underpins the immune response to a variety of infections, is immunometabolism, namely glycolysis and oxidative phosphorylation (OXPHOS). We hypothesise that in addition to their direct bactericidal effect on Mycobacterium tuberculosis (Mtb), current TB antimicrobials can modulate immunometabolic profiles and alter mitochondrial function in primary human macrophages. Human monocyte-derived macrophages (hMDMs) were differentiated from PBMCs isolated from healthy blood donors, and treated with four first-line and six second-line TB antimicrobials three hours post stimulation with either iH37Rv-Mtb or lipopolysaccharide (LPS). 24 h post stimulation, baseline metabolism and mitochondrial function were determined using the Seahorse Extracellular Flux Analyser. The effect of these antimicrobials on cytokine and chemokine production was also assayed using Meso Scale Discovery Multi-Array technology. We show that some of the TB antimicrobials tested can significantly alter OXPHOS and glycolysis in uninfected, iH37Rv-Mtb, and LPS-stimulated hMDMs. We also demonstrate how these antimicrobial-induced immunometabolic effects are linked with alterations in mitochondrial function. Our results show that TB antimicrobials, specifically clofazimine, can modify host immunometabolism and mitochondrial function. Moreover, clofazimine significantly increased the production of IL-6 in human macrophages that were stimulated with iH37Rv-Mtb. This provides further insight into the use of some of these TB antimicrobials as potential host-directed therapies in patients with early and active disease, which could help to inform TB treatment strategies in the future.

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Section: Tb Antimicrobials Differentially Affect Glycolysis and Oxidative Phosphorylation Levels Inmentioning

confidence: 99%

Section: Tb Antimicrobials Differentially Affect Glycolysis and Oxidative Phosphorylation Levels Inmentioning

confidence: 99%

The Effect of Tuberculosis Antimicrobials on the Immunometabolic Profiles of Primary Human Macrophages Stimulated with Mycobacterium tuberculosis

Cahill

Cox

O’Connell

et al. 2021

IJMS

View full text Add to dashboard Cite

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Comparison of the XTT and resazurin assays for quantification of the metabolic activity of Staphylococcus aureus biofilm

Alonso

Cruces

Pérez

et al. 2017

Journal of Microbiological Methods

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Modeling Tubercular ESX-1 Secretion Using Mycobacterium marinum

Chirakos

Balaram

Conrad

et al. 2020

Microbiol Mol Biol Rev

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SUMMARY Pathogenic mycobacteria cause chronic and acute diseases ranging from human tuberculosis (TB) to nontubercular infections. Mycobacterium tuberculosis causes both acute and chronic human tuberculosis. Environmentally acquired nontubercular mycobacteria (NTM) cause chronic disease in humans and animals. Not surprisingly, NTM and M. tuberculosis often use shared molecular mechanisms to survive within the host. The ESX-1 system is a specialized secretion system that is essential for virulence and is functionally conserved between M. tuberculosis and Mycobacterium marinum. M. marinum is an NTM found in both salt water and freshwater that is often used to study mycobacterial virulence. Since the discovery of the secretion system in 2003, the use of both M. tuberculosis and M. marinum has defined the conserved molecular mechanisms underlying protein secretion and the lytic and regulatory activities of the ESX-1 system. Here, we review the trajectory of the field, including key discoveries regarding the ESX-1 system. We highlight the contributions of M. marinum studies and the conserved and unique aspects of the ESX-1 secretion system.

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Pyrazinamide susceptibility testing in <I>Mycobacterium tuberculosis</I> using the fast resazurin microtiter assay plate

Cited by 3 publications

References 0 publications

The Effect of Tuberculosis Antimicrobials on the Immunometabolic Profiles of Primary Human Macrophages Stimulated with Mycobacterium tuberculosis

The Effect of Tuberculosis Antimicrobials on the Immunometabolic Profiles of Primary Human Macrophages Stimulated with Mycobacterium tuberculosis

Comparison of the XTT and resazurin assays for quantification of the metabolic activity of Staphylococcus aureus biofilm

Modeling Tubercular ESX-1 Secretion Using Mycobacterium marinum

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