Limits...
Pro-atherogenic lung and oral pathogens induce an inflammatory response in human and mouse mast cells.

Oksaharju A, Lappalainen J, Tuomainen AM, Pussinen PJ, Puolakkainen M, Kovanen PT, Lindstedt KA - J. Cell. Mol. Med. (2009)

Bottom Line: Human monocyte-derived macrophages that were pre-infected with Cpn also induced a significant pro-inflammatory response in human mast cells, both in cocultures and when preconditioned media from Cpn-infected macrophages were used.Intranasal and intravenous administration of live Cpn and Aa, respectively induced an accumulation of activated mast cells in the aortic sinus of apolipoprotein E-deficient mice, however, with varying responses in the systemic levels of lipopolysaccharide (LPS) and TNF-alpha.Pro-atherogenic Cpn and Aa induce a pro-inflammatory response in cultured human connective tissue-type mast cells and activation of mouse aortic mast cells in vivo.

View Article: PubMed Central - PubMed

Affiliation: Wihuri Research Institute, Kalliolinantie, Helsinki, Finland.

ABSTRACT
A broad variety of microbes are present in atherosclerotic plaques and chronic bacterial infection increases the risk of atherosclerosis by mechanisms that have remained vague. One possible mechanism is that bacteria or bacterial products activate plaque mast cells that are known to participate in the pathogenesis of atherosclerosis. Here, we show by real-time PCR analysis and ELISA that Chlamydia pneumoniae (Cpn) and a periodontal pathogen, Aggregatibacter actinomycetemcomitans (Aa), both induce a time and concentration-dependent expression and secretion of interleukin 8 (IL-8), tumour necrosis factor-alpha (TNF-alpha) and monocyte chemoattractant protein-1 (MCP-1) by cultured human peripheral blood-derived mast cells, but not anti-inflammatory molecules, such as IL-10 or transforming growth factor beta 1 (TGF-beta 1). The IL-8 and MCP-1 responses were immediate, whereas the onset of TNF-alpha secretion was delayed. The Cpn-mediated pro-inflammatory effect was attenuated when the bacteria were inactivated by UV-treatment. Human monocyte-derived macrophages that were pre-infected with Cpn also induced a significant pro-inflammatory response in human mast cells, both in cocultures and when preconditioned media from Cpn-infected macrophages were used. Intranasal and intravenous administration of live Cpn and Aa, respectively induced an accumulation of activated mast cells in the aortic sinus of apolipoprotein E-deficient mice, however, with varying responses in the systemic levels of lipopolysaccharide (LPS) and TNF-alpha. Pro-atherogenic Cpn and Aa induce a pro-inflammatory response in cultured human connective tissue-type mast cells and activation of mouse aortic mast cells in vivo.

Show MeSH

Related in: MedlinePlus

Temporal induction of pro-inflammatory cytokines and chemokines. Cultured human mast cells were infected with Cpn in a cell-to-bacteria ratio of 1: 10 and incubated for 0, 1, 3, 6 and 24 hrs. Quantitative analysis of interleukin 8 (IL-8) (A), MCP-1 (B) and tumour necrosis factor-α (TNF-a) (C) mRNA expression was performed with real-time PCR and the concentrations of these cytokines in the incubation media were detected by ELISA (IL-8 [D], MCP-1 [E] and TNF-α[F]). Data are means ± SD and the results shown are representative of four independent experiments.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3823039&req=5

fig02: Temporal induction of pro-inflammatory cytokines and chemokines. Cultured human mast cells were infected with Cpn in a cell-to-bacteria ratio of 1: 10 and incubated for 0, 1, 3, 6 and 24 hrs. Quantitative analysis of interleukin 8 (IL-8) (A), MCP-1 (B) and tumour necrosis factor-α (TNF-a) (C) mRNA expression was performed with real-time PCR and the concentrations of these cytokines in the incubation media were detected by ELISA (IL-8 [D], MCP-1 [E] and TNF-α[F]). Data are means ± SD and the results shown are representative of four independent experiments.

Mentions: To test the ability of infectious pro-atherogenic pathogens, Cpn and two strains (AT445b and JP2) of Aa, to induce an inflammatory response in cultured human mast cells, the mast cells were infected with the different bacteria for 6 hrs at 37°C in 5% CO2, after which two selected atherosclerosis-related inflammatory cytokines (TNF-α and IL-8) and one chemokine (MCP-1) were analysed. Cpn induced a significant expression of both IL-8 (P= 0.00007) and TNF-α (P= 0.0026) mRNA, whereas the expression of MCP-1 mRNA was only slightly increased (statistically not significant) (Fig. 1). Of the two Aa strains, only the JP2 strain induced a significant expression of IL-8 (P= 0.0039) mRNA after 6 hrs of incubation (Fig. 1A), whereas the expression of TNF-α and MCP-1 mRNA was only slightly increased (statistically not significant). The AT445b strain did not induce a statistically significant mRNA expression of the tested cytokines after 6 hrs of incubation. Following inoculation with Cpn, IL-8 and MCP-1 mRNA rapidly increased (within 1 hr), and also rapidly returned to the basal level (Fig. 2, left side). In sharp contrast to this rapid fluctuation, the expression of TNF-a mRNA was increased only after 3 hrs of bacterial inoculation, and was further increased at 24 hrs of incubation.


Pro-atherogenic lung and oral pathogens induce an inflammatory response in human and mouse mast cells.

Oksaharju A, Lappalainen J, Tuomainen AM, Pussinen PJ, Puolakkainen M, Kovanen PT, Lindstedt KA - J. Cell. Mol. Med. (2009)

Temporal induction of pro-inflammatory cytokines and chemokines. Cultured human mast cells were infected with Cpn in a cell-to-bacteria ratio of 1: 10 and incubated for 0, 1, 3, 6 and 24 hrs. Quantitative analysis of interleukin 8 (IL-8) (A), MCP-1 (B) and tumour necrosis factor-α (TNF-a) (C) mRNA expression was performed with real-time PCR and the concentrations of these cytokines in the incubation media were detected by ELISA (IL-8 [D], MCP-1 [E] and TNF-α[F]). Data are means ± SD and the results shown are representative of four independent experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3823039&req=5

fig02: Temporal induction of pro-inflammatory cytokines and chemokines. Cultured human mast cells were infected with Cpn in a cell-to-bacteria ratio of 1: 10 and incubated for 0, 1, 3, 6 and 24 hrs. Quantitative analysis of interleukin 8 (IL-8) (A), MCP-1 (B) and tumour necrosis factor-α (TNF-a) (C) mRNA expression was performed with real-time PCR and the concentrations of these cytokines in the incubation media were detected by ELISA (IL-8 [D], MCP-1 [E] and TNF-α[F]). Data are means ± SD and the results shown are representative of four independent experiments.
Mentions: To test the ability of infectious pro-atherogenic pathogens, Cpn and two strains (AT445b and JP2) of Aa, to induce an inflammatory response in cultured human mast cells, the mast cells were infected with the different bacteria for 6 hrs at 37°C in 5% CO2, after which two selected atherosclerosis-related inflammatory cytokines (TNF-α and IL-8) and one chemokine (MCP-1) were analysed. Cpn induced a significant expression of both IL-8 (P= 0.00007) and TNF-α (P= 0.0026) mRNA, whereas the expression of MCP-1 mRNA was only slightly increased (statistically not significant) (Fig. 1). Of the two Aa strains, only the JP2 strain induced a significant expression of IL-8 (P= 0.0039) mRNA after 6 hrs of incubation (Fig. 1A), whereas the expression of TNF-α and MCP-1 mRNA was only slightly increased (statistically not significant). The AT445b strain did not induce a statistically significant mRNA expression of the tested cytokines after 6 hrs of incubation. Following inoculation with Cpn, IL-8 and MCP-1 mRNA rapidly increased (within 1 hr), and also rapidly returned to the basal level (Fig. 2, left side). In sharp contrast to this rapid fluctuation, the expression of TNF-a mRNA was increased only after 3 hrs of bacterial inoculation, and was further increased at 24 hrs of incubation.

Bottom Line: Human monocyte-derived macrophages that were pre-infected with Cpn also induced a significant pro-inflammatory response in human mast cells, both in cocultures and when preconditioned media from Cpn-infected macrophages were used.Intranasal and intravenous administration of live Cpn and Aa, respectively induced an accumulation of activated mast cells in the aortic sinus of apolipoprotein E-deficient mice, however, with varying responses in the systemic levels of lipopolysaccharide (LPS) and TNF-alpha.Pro-atherogenic Cpn and Aa induce a pro-inflammatory response in cultured human connective tissue-type mast cells and activation of mouse aortic mast cells in vivo.

View Article: PubMed Central - PubMed

Affiliation: Wihuri Research Institute, Kalliolinantie, Helsinki, Finland.

ABSTRACT
A broad variety of microbes are present in atherosclerotic plaques and chronic bacterial infection increases the risk of atherosclerosis by mechanisms that have remained vague. One possible mechanism is that bacteria or bacterial products activate plaque mast cells that are known to participate in the pathogenesis of atherosclerosis. Here, we show by real-time PCR analysis and ELISA that Chlamydia pneumoniae (Cpn) and a periodontal pathogen, Aggregatibacter actinomycetemcomitans (Aa), both induce a time and concentration-dependent expression and secretion of interleukin 8 (IL-8), tumour necrosis factor-alpha (TNF-alpha) and monocyte chemoattractant protein-1 (MCP-1) by cultured human peripheral blood-derived mast cells, but not anti-inflammatory molecules, such as IL-10 or transforming growth factor beta 1 (TGF-beta 1). The IL-8 and MCP-1 responses were immediate, whereas the onset of TNF-alpha secretion was delayed. The Cpn-mediated pro-inflammatory effect was attenuated when the bacteria were inactivated by UV-treatment. Human monocyte-derived macrophages that were pre-infected with Cpn also induced a significant pro-inflammatory response in human mast cells, both in cocultures and when preconditioned media from Cpn-infected macrophages were used. Intranasal and intravenous administration of live Cpn and Aa, respectively induced an accumulation of activated mast cells in the aortic sinus of apolipoprotein E-deficient mice, however, with varying responses in the systemic levels of lipopolysaccharide (LPS) and TNF-alpha. Pro-atherogenic Cpn and Aa induce a pro-inflammatory response in cultured human connective tissue-type mast cells and activation of mouse aortic mast cells in vivo.

Show MeSH
Related in: MedlinePlus