Mycobacterial <i>para</i>-Hydroxybenzoic Acid-Derivatives (<i>p</i>HBADs) and Related Structures Induce Macrophage Innate Memory

Lundahl, Mimmi L. E.; Lynch, Dylan M.; Barnes, Danielle; McSweeney, Lauren; Gorman, Ashleigh; Lebre, Filipa; Gordon, Stephen V.; Lavelle, Ed C.; Scanlan, Eoin M.

doi:10.1021/acschembio.0c00378

Cited by 5 publications

(5 citation statements)

References 34 publications

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“…Previous work has shown that exogenous radiolabeled p HB is taken up by mycobacterial cells and metabolically incorporated into cell–surface PGL . While p HB is also used in the biosynthesis of other p -hydroxybenzoic acid derivatives ( p HBADs), − these metabolites are not associated with the cell envelopethey are either cytosolic or secretedand therefore would not be expected to confound the visualization of membrane-associated PGL. We synthesized both 2- and 3-azido p HB as described in the Supporting Information.…”

Section: Results and Discussionmentioning

confidence: 99%

Bioorthogonal Metabolic Labeling of the Virulence Factor Phenolic Glycolipid in Mycobacteria

Guzmán,

Cambier,

Cheng

et al. 2024

ACS Chem. Biol.

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Surface lipids on pathogenic mycobacteria modulate infection outcomes by regulating host immune responses. Phenolic glycolipid (PGL) is a host-modulating surface lipid that varies among clinical Mycobacterium tuberculosis strains. PGL is also found in Mycobacterium marinum, where it promotes infection of zebrafish through effects on the innate immune system. Given the important role this lipid plays in the host−pathogen relationship, tools for profiling its abundance, spatial distribution, and dynamics are needed. Here, we report a strategy for imaging PGL in live mycobacteria using bioorthogonal metabolic labeling. We functionalized the PGL precursor p-hydroxybenzoic acid (pHB) with an azide group (3-azido pHB). When fed to mycobacteria, 3-azido pHB was incorporated into the cell surface, which could then be visualized via the bioorthogonal conjugation of a fluorescent probe. We confirmed that 3-azido pHB incorporates into PGL using mass spectrometry methods and demonstrated selectivity for PGL-producing M. marinum and M. tuberculosis strains. Finally, we applied this metabolic labeling strategy to study the dynamics of PGL within the mycobacterial membrane. This new tool enables visualization of PGL that may facilitate studies of mycobacterial pathogenesis.

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Section: Results and Discussionmentioning

confidence: 99%

Bioorthogonal Metabolic Labeling of the Virulence Factor Phenolic Glycolipid in Mycobacteria

Guzmán,

Cambier,

Cheng

et al. 2024

ACS Chem. Biol.

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“…Previous work has shown that exogenous radiolabeled pHB is taken up by mycobacterial cells and metabolically incorporated into cell-surface PGL. 8 While pHB is Phenolphthiocerol, phenolphthiodiolone, or phenolphthiotriol also used in the biosynthesis of other p-hydroxybenzoic acid derivatives (pHBADs), [36][37][38][39] these metabolites are not associated with the cell envelope -they are either cytosolic or secreted -and therefore would not be expected to confound the visualization of membrane-associated PGL. We synthesized both 2-and 3-azido pHB as described in the supporting information.…”

Section: Resultsmentioning

confidence: 99%

Bioorthogonal metabolic labeling of the virulence factor phenolic glycolipid in mycobacteria

Guzmán,

Cambier,

Cheng

et al. 2023

Preprint

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Surface lipids on pathogenic mycobacteria modulate infection outcomes by regulating host immune responses. Phenolic glycolipid (PGL) is a host-modulating surface lipid that varies among clinicalMycobacterium tuberculosisstrains. PGL is also found inMycobacterium marinumwhere it promotes infection of zebrafish through effects on the innate immune system. Given the important role this lipid plays in the host-pathogen relationship, tools for profiling its abundance, spatial distribution, and dynamics are needed. Here we report a strategy for imaging PGL in live mycobacteria using bioorthogonal metabolic labeling. We functionalized the PGL precursorp-hydroxybenzoic acid (pHB) with an azide group (3-azidopHB). When fed to mycobacteria, 3-azidopHB was incorporated into the cell surface, which could then be visualized via bioorthogonal conjugation of a fluorescent probe. We confirmed that 3-azidopHB incorporates into PGL using mass spectrometry methods and demonstrated selectivity for PGL-producingMycobacterium marinumandMycobacterium tuberculosisstrains. Finally, we applied this metabolic labeling strategy to study the dynamics of PGL within the mycobacterial membrane. This new tool enables visualization of PGL which may facilitate studies of mycobacterial pathogenesis.

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“…Conversely, there is a risk that certain immune challenges could lead to innate immune re-programming that hinders protective immune responses. For instance, recent reports have demonstrated how virulent M. tuberculosis ( Khan et al, 2020 ) and mycobacterial phenolic glycans ( Lundahl et al, 2020 ) can program macrophages to attenuate bactericidal responses to subsequent mycobacterial challenge. In the current study, macrophage activation caused by former or concurrent parasitic infections is considered regarding the possibility of innate immune re-programming that hinders protective immune responses against this disease.…”

Section: Introductionmentioning

confidence: 99%

Macrophage innate training induced by IL-4 and IL-13 activation enhances OXPHOS driven anti-mycobacterial responses

Lundahl

Mitermite

Ryan

et al. 2022

eLife

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Macrophages are a highly adaptive population of innate immune cells. Polarization with IFNγ and LPS into the 'classically activated' M1 macrophage enhances pro-inflammatory and microbicidal responses, important for eradicating bacteria such as Mycobacterium tuberculosis. By contrast, 'alternatively activated' M2 macrophages, polarized with IL-4, oppose bactericidal mechanisms and allow mycobacterial growth. These activation states are accompanied by distinct metabolic profiles, where M1 macrophages favor near exclusive use of glycolysis, whereas M2 macrophages up-regulate oxidative phosphorylation (OXPHOS). Here we demonstrate that activation with IL-4 and IL-13 counterintuitively induces protective innate memory against mycobacterial challenge. In human and murine models, prior activation with IL-4/13 enhances pro-inflammatory cytokine secretion in response to a secondary stimulation with mycobacterial ligands. In our murine model, enhanced killing capacity is also demonstrated. Despite this switch in phenotype, IL-4/13 trained murine macrophages do not demonstrate M1-typical metabolism, instead retaining heightened use of OXPHOS. Moreover, inhibition of OXPHOS with oligomycin, 2-deoxy glucose or BPTES all impeded heightened pro-inflammatory cytokine responses from IL-4/13 trained macrophages. Lastly, this work identifies that IL-10 attenuates protective IL-4/13 training, impeding pro-inflammatory and bactericidal mechanisms. In summary, this work provides new and unexpected insight into alternative macrophage activation states in the context of mycobacterial infection.

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Mycobacterial para-Hydroxybenzoic Acid-Derivatives (pHBADs) and Related Structures Induce Macrophage Innate Memory

Cited by 5 publications

References 34 publications

Bioorthogonal Metabolic Labeling of the Virulence Factor Phenolic Glycolipid in Mycobacteria

Bioorthogonal Metabolic Labeling of the Virulence Factor Phenolic Glycolipid in Mycobacteria

Bioorthogonal metabolic labeling of the virulence factor phenolic glycolipid in mycobacteria

Macrophage innate training induced by IL-4 and IL-13 activation enhances OXPHOS driven anti-mycobacterial responses

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