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ESX1-dependent fractalkine mediates chemotaxis and Mycobacterium tuberculosis infection in humans.

Hingley-Wilson SM, Connell D, Pollock K, Hsu T, Tchilian E, Sykes A, Grass L, Potiphar L, Bremang S, Kon OM, Jacobs WR, Lalvani A - Tuberculosis (Edinb) (2014)

Bottom Line: The M. tuberculosis ESX1 mutant, which has a non-functional type VII secretion system, induced significantly less production of the host macrophage-derived chemokine fractalkine (CX3CL1).Fractalkine levels were raised in vivo at tuberculous disease sites in humans and were significantly associated with increased CD11b+ monocytic cellular recruitment and extent of granulomatous disease.These findings suggest a novel fractalkine-dependent ESX1-mediated mechanism in early tuberculous disease pathogenesis in humans.

View Article: PubMed Central - PubMed

Affiliation: Tuberculosis Research Centre, Department of Respiratory Medicine, National Heart and Lung Institute, Imperial College London, London W2 1PG, United Kingdom. Electronic address: s.hingley-wilson@surrey.ac.uk.

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MMP-9 is produced by human macrophages at 24 h post-infection, but is not ESX1-mediated. Cells were infected at an M.O.I. of 10:1 or exposed to conditioned medium for 24 h. U = uninfected, Rv = Mtb H37Rv, ΔESX1 = Mtb ESX1 mutant, C = complemented ESX1 mutant, P = Mtb plus piceatannol, Ab = Mtb plus anti-fractalkine antibody, coRv = conditioned medium from Rv infected cells, coΔ = conditioned medium from ESX-1 infected cells, LPS = 1 ng/ml a) THP-1 cells, b) THP-1 cells plus overnight 10 U IFN-γ, c)MDMs, d) A549 alveolar epithelial cells. Data is representative of two replicates and 2 independent experiments and shows standard error bars.
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fig3: MMP-9 is produced by human macrophages at 24 h post-infection, but is not ESX1-mediated. Cells were infected at an M.O.I. of 10:1 or exposed to conditioned medium for 24 h. U = uninfected, Rv = Mtb H37Rv, ΔESX1 = Mtb ESX1 mutant, C = complemented ESX1 mutant, P = Mtb plus piceatannol, Ab = Mtb plus anti-fractalkine antibody, coRv = conditioned medium from Rv infected cells, coΔ = conditioned medium from ESX-1 infected cells, LPS = 1 ng/ml a) THP-1 cells, b) THP-1 cells plus overnight 10 U IFN-γ, c)MDMs, d) A549 alveolar epithelial cells. Data is representative of two replicates and 2 independent experiments and shows standard error bars.

Mentions: Elucidation of the fundamental role of M. marinum-induced ESX-1-dependent MMP-9 secretion in zebrafish embryos [4] after we generated the above data prompted us to investigate ESX1-dependence of MMP-9 secretion in our human system. We assessed production of MMP-9 in THP-1 cells, both with and without IFN-γ, and in human MDMs and found no difference between Mtb H37Rv and the Mtb ΔESX1 mutant. This was also the case with cells activated with conditioned media from infected macrophages (Figure 3a–c). Although we observed substantial production of MMP9 in human alveolar epithelial cells (A549 cell line; Figure 3d) there was no ESX-1 mediated difference. The zebrafish immune system differs substantially from the human, so we studied several other MMPs (1, 2, 7 and 10) to see if an alternative MMP might be acting in the place of the zebrafish MMP9 and there was no observable ESX1-mediated difference in the MMPs 1, 2 7 or 10 (Supplementary Figure 1).


ESX1-dependent fractalkine mediates chemotaxis and Mycobacterium tuberculosis infection in humans.

Hingley-Wilson SM, Connell D, Pollock K, Hsu T, Tchilian E, Sykes A, Grass L, Potiphar L, Bremang S, Kon OM, Jacobs WR, Lalvani A - Tuberculosis (Edinb) (2014)

MMP-9 is produced by human macrophages at 24 h post-infection, but is not ESX1-mediated. Cells were infected at an M.O.I. of 10:1 or exposed to conditioned medium for 24 h. U = uninfected, Rv = Mtb H37Rv, ΔESX1 = Mtb ESX1 mutant, C = complemented ESX1 mutant, P = Mtb plus piceatannol, Ab = Mtb plus anti-fractalkine antibody, coRv = conditioned medium from Rv infected cells, coΔ = conditioned medium from ESX-1 infected cells, LPS = 1 ng/ml a) THP-1 cells, b) THP-1 cells plus overnight 10 U IFN-γ, c)MDMs, d) A549 alveolar epithelial cells. Data is representative of two replicates and 2 independent experiments and shows standard error bars.
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Related In: Results  -  Collection

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fig3: MMP-9 is produced by human macrophages at 24 h post-infection, but is not ESX1-mediated. Cells were infected at an M.O.I. of 10:1 or exposed to conditioned medium for 24 h. U = uninfected, Rv = Mtb H37Rv, ΔESX1 = Mtb ESX1 mutant, C = complemented ESX1 mutant, P = Mtb plus piceatannol, Ab = Mtb plus anti-fractalkine antibody, coRv = conditioned medium from Rv infected cells, coΔ = conditioned medium from ESX-1 infected cells, LPS = 1 ng/ml a) THP-1 cells, b) THP-1 cells plus overnight 10 U IFN-γ, c)MDMs, d) A549 alveolar epithelial cells. Data is representative of two replicates and 2 independent experiments and shows standard error bars.
Mentions: Elucidation of the fundamental role of M. marinum-induced ESX-1-dependent MMP-9 secretion in zebrafish embryos [4] after we generated the above data prompted us to investigate ESX1-dependence of MMP-9 secretion in our human system. We assessed production of MMP-9 in THP-1 cells, both with and without IFN-γ, and in human MDMs and found no difference between Mtb H37Rv and the Mtb ΔESX1 mutant. This was also the case with cells activated with conditioned media from infected macrophages (Figure 3a–c). Although we observed substantial production of MMP9 in human alveolar epithelial cells (A549 cell line; Figure 3d) there was no ESX-1 mediated difference. The zebrafish immune system differs substantially from the human, so we studied several other MMPs (1, 2, 7 and 10) to see if an alternative MMP might be acting in the place of the zebrafish MMP9 and there was no observable ESX1-mediated difference in the MMPs 1, 2 7 or 10 (Supplementary Figure 1).

Bottom Line: The M. tuberculosis ESX1 mutant, which has a non-functional type VII secretion system, induced significantly less production of the host macrophage-derived chemokine fractalkine (CX3CL1).Fractalkine levels were raised in vivo at tuberculous disease sites in humans and were significantly associated with increased CD11b+ monocytic cellular recruitment and extent of granulomatous disease.These findings suggest a novel fractalkine-dependent ESX1-mediated mechanism in early tuberculous disease pathogenesis in humans.

View Article: PubMed Central - PubMed

Affiliation: Tuberculosis Research Centre, Department of Respiratory Medicine, National Heart and Lung Institute, Imperial College London, London W2 1PG, United Kingdom. Electronic address: s.hingley-wilson@surrey.ac.uk.

Show MeSH
Related in: MedlinePlus