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Host targeted activity of pyrazinamide in Mycobacterium tuberculosis infection.

Manca C, Koo MS, Peixoto B, Fallows D, Kaplan G, Subbian S - PLoS ONE (2013)

Bottom Line: In vitro we have examined the effect of PZA on cytokine and chemokine release by Mtb-infected or Toll-like receptor (TLR) -stimulated primary human monocytes.Here, we report that PZA treatment of Mtb-infected human monocytes and mice significantly reduces the release of pro-inflammatory cytokines and chemokines, including IL-1β, IL-6, TNF-α and MCP-1 at the protein and at the gene transcription levels, respectively.In addition, our results suggest that inclusion or exclusion of PZA in the TB treatment regimen could potentially affect the biomarker signature detected in the circulation of TB patients.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Mycobacterial Immunity and Pathogenesis, Public Health Research Institute (PHRI), New Jersey Medical School, Rutgers Biomedical and Health Sciences, Rutgers The State University of New Jersey, Newark, New Jersey, United States of America.

ABSTRACT
Pyrazinamide (PZA) is one of the first line antibiotics used for the treatment of tuberculosis (TB). In the present study, we have used in vitro and in vivo systems to investigate whether PZA, in addition to its known anti-mycobacterial properties, modulate the host immune response during Mycobacterium tuberculosis (Mtb) infection. In vitro we have examined the effect of PZA on cytokine and chemokine release by Mtb-infected or Toll-like receptor (TLR) -stimulated primary human monocytes. In vivo, we have investigated at the transcriptional levels using genome-wide microarray gene expression analysis, whether PZA treatment of Mtb-infected mice alters the host immune response to Mtb infection in the lungs. Here, we report that PZA treatment of Mtb-infected human monocytes and mice significantly reduces the release of pro-inflammatory cytokines and chemokines, including IL-1β, IL-6, TNF-α and MCP-1 at the protein and at the gene transcription levels, respectively. Data from microarray analysis also reveal that PZA treatment of Mtb-infected mice significantly alters the expression level of genes involved in the regulation of the pro-inflammatory mediators, lung inflammatory response and TLR signaling networks. Specifically, genes coding for adenylate cyclase and Peroxisome-Proliferator Activated Receptor (PPAR), molecules known for their anti-inflammatory effect, were found to be up-regulated in the lungs of PZA-treated Mtb-infected mice. Based on the microarray findings, we propose that PZA treatment modulates the host immune response to Mtb infection by reducing pro-inflammatory cytokine production, probably through PPAR- and NF-kB- dependent pathways. In addition, our results suggest that inclusion or exclusion of PZA in the TB treatment regimen could potentially affect the biomarker signature detected in the circulation of TB patients.

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PZA effect on monocytes stimulated with selected TLR agonists. Human monocytes treated with PZA (10 or 50 µg/ml) were simultaneously stimulated with (A) TLR4 (LPS, 100 ng/ml) or (B) TLR2/6 (Pam2, 250 ng/ml) and TLR2/1 (Pam3, 250 ng/ml) agonists. Pro-inflammatory and down-regulatory mediators were measured in the culture supernatants at 24 hours post-stimulation. Data are from 4 – 7 independent experiments (independent donors; N =  4 – 7) performed in duplicate and presented as percentage induction relative to PZA-untreated TLR-stimulated cells ± SD. * statistically significant; P ≤ 0.05 compared with the PZA-untreated TLR-stimulated cells.
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pone-0074082-g002: PZA effect on monocytes stimulated with selected TLR agonists. Human monocytes treated with PZA (10 or 50 µg/ml) were simultaneously stimulated with (A) TLR4 (LPS, 100 ng/ml) or (B) TLR2/6 (Pam2, 250 ng/ml) and TLR2/1 (Pam3, 250 ng/ml) agonists. Pro-inflammatory and down-regulatory mediators were measured in the culture supernatants at 24 hours post-stimulation. Data are from 4 – 7 independent experiments (independent donors; N =  4 – 7) performed in duplicate and presented as percentage induction relative to PZA-untreated TLR-stimulated cells ± SD. * statistically significant; P ≤ 0.05 compared with the PZA-untreated TLR-stimulated cells.

Mentions: To confirm that PZA can affect cytokine/chemokine production by monocytes independent of changes in the Mtb bacillary load monocytes were stimulated with TLR agonists and the impact of simultaneous treatment with PZA on the release of specific cytokines/chemokines was evaluated and found to differ among the agonists. When LPS (TLR4 agonist) was used to stimulate monocytes, PZA treatment showed a concentration-dependent inhibitory effect in the release of all four pro-inflammatory mediators tested, and did not alter the release of IL-1Ra, a known IL-1β inhibitor, in Mtb-infected cells (Figure 2A). Our observation that the levels of IL-1Ra were unaffected by PZA treatment suggested that the reduction in the levels of IL-1β was not IL-1Ra-dependent. When PZA-treated cells were simultaneously stimulated with Pam2CSK4 (TLR2/6) or Pam3CSK4 (TLR2/1) agonists and PZA, concentration-dependent inhibition was seen only for IL-1β or MCP-1, respectively (Figure 2B). Thus, the TLR stimulation data confirmed that PZA affected monocyte cytokine/chemokine production independently of Mtb infection or Mtb bacillary load in the monocytes.


Host targeted activity of pyrazinamide in Mycobacterium tuberculosis infection.

Manca C, Koo MS, Peixoto B, Fallows D, Kaplan G, Subbian S - PLoS ONE (2013)

PZA effect on monocytes stimulated with selected TLR agonists. Human monocytes treated with PZA (10 or 50 µg/ml) were simultaneously stimulated with (A) TLR4 (LPS, 100 ng/ml) or (B) TLR2/6 (Pam2, 250 ng/ml) and TLR2/1 (Pam3, 250 ng/ml) agonists. Pro-inflammatory and down-regulatory mediators were measured in the culture supernatants at 24 hours post-stimulation. Data are from 4 – 7 independent experiments (independent donors; N =  4 – 7) performed in duplicate and presented as percentage induction relative to PZA-untreated TLR-stimulated cells ± SD. * statistically significant; P ≤ 0.05 compared with the PZA-untreated TLR-stimulated cells.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3755974&req=5

pone-0074082-g002: PZA effect on monocytes stimulated with selected TLR agonists. Human monocytes treated with PZA (10 or 50 µg/ml) were simultaneously stimulated with (A) TLR4 (LPS, 100 ng/ml) or (B) TLR2/6 (Pam2, 250 ng/ml) and TLR2/1 (Pam3, 250 ng/ml) agonists. Pro-inflammatory and down-regulatory mediators were measured in the culture supernatants at 24 hours post-stimulation. Data are from 4 – 7 independent experiments (independent donors; N =  4 – 7) performed in duplicate and presented as percentage induction relative to PZA-untreated TLR-stimulated cells ± SD. * statistically significant; P ≤ 0.05 compared with the PZA-untreated TLR-stimulated cells.
Mentions: To confirm that PZA can affect cytokine/chemokine production by monocytes independent of changes in the Mtb bacillary load monocytes were stimulated with TLR agonists and the impact of simultaneous treatment with PZA on the release of specific cytokines/chemokines was evaluated and found to differ among the agonists. When LPS (TLR4 agonist) was used to stimulate monocytes, PZA treatment showed a concentration-dependent inhibitory effect in the release of all four pro-inflammatory mediators tested, and did not alter the release of IL-1Ra, a known IL-1β inhibitor, in Mtb-infected cells (Figure 2A). Our observation that the levels of IL-1Ra were unaffected by PZA treatment suggested that the reduction in the levels of IL-1β was not IL-1Ra-dependent. When PZA-treated cells were simultaneously stimulated with Pam2CSK4 (TLR2/6) or Pam3CSK4 (TLR2/1) agonists and PZA, concentration-dependent inhibition was seen only for IL-1β or MCP-1, respectively (Figure 2B). Thus, the TLR stimulation data confirmed that PZA affected monocyte cytokine/chemokine production independently of Mtb infection or Mtb bacillary load in the monocytes.

Bottom Line: In vitro we have examined the effect of PZA on cytokine and chemokine release by Mtb-infected or Toll-like receptor (TLR) -stimulated primary human monocytes.Here, we report that PZA treatment of Mtb-infected human monocytes and mice significantly reduces the release of pro-inflammatory cytokines and chemokines, including IL-1β, IL-6, TNF-α and MCP-1 at the protein and at the gene transcription levels, respectively.In addition, our results suggest that inclusion or exclusion of PZA in the TB treatment regimen could potentially affect the biomarker signature detected in the circulation of TB patients.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Mycobacterial Immunity and Pathogenesis, Public Health Research Institute (PHRI), New Jersey Medical School, Rutgers Biomedical and Health Sciences, Rutgers The State University of New Jersey, Newark, New Jersey, United States of America.

ABSTRACT
Pyrazinamide (PZA) is one of the first line antibiotics used for the treatment of tuberculosis (TB). In the present study, we have used in vitro and in vivo systems to investigate whether PZA, in addition to its known anti-mycobacterial properties, modulate the host immune response during Mycobacterium tuberculosis (Mtb) infection. In vitro we have examined the effect of PZA on cytokine and chemokine release by Mtb-infected or Toll-like receptor (TLR) -stimulated primary human monocytes. In vivo, we have investigated at the transcriptional levels using genome-wide microarray gene expression analysis, whether PZA treatment of Mtb-infected mice alters the host immune response to Mtb infection in the lungs. Here, we report that PZA treatment of Mtb-infected human monocytes and mice significantly reduces the release of pro-inflammatory cytokines and chemokines, including IL-1β, IL-6, TNF-α and MCP-1 at the protein and at the gene transcription levels, respectively. Data from microarray analysis also reveal that PZA treatment of Mtb-infected mice significantly alters the expression level of genes involved in the regulation of the pro-inflammatory mediators, lung inflammatory response and TLR signaling networks. Specifically, genes coding for adenylate cyclase and Peroxisome-Proliferator Activated Receptor (PPAR), molecules known for their anti-inflammatory effect, were found to be up-regulated in the lungs of PZA-treated Mtb-infected mice. Based on the microarray findings, we propose that PZA treatment modulates the host immune response to Mtb infection by reducing pro-inflammatory cytokine production, probably through PPAR- and NF-kB- dependent pathways. In addition, our results suggest that inclusion or exclusion of PZA in the TB treatment regimen could potentially affect the biomarker signature detected in the circulation of TB patients.

Show MeSH
Related in: MedlinePlus