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Subtoxic Doses of Cadmium Modulate Inflammatory Properties of Murine RAW 264.7 Macrophages.

Riemschneider S, Herzberg M, Lehmann J - Biomed Res Int (2015)

Bottom Line: Cadmium alone caused a dose-dependent decreased viability of exposed cells.Also production of antibacterial NO was significantly reduced through exposure to 10 µM Cd, maybe explaining better survival of macrophages.Additionally, we could show by analysis via ICP-MS that different effects of Cd in nonactivated and activated macrophages definitely did not result from different Cd uptake rates.

View Article: PubMed Central - PubMed

Affiliation: Fraunhofer Institute for Cell Therapy and Immunology (IZI), 04103 Leipzig, Germany.

ABSTRACT
Cadmium (Cd) is a toxic heavy metal that exhibits various adverse effects in the human and animal organism. Its resemblance to essential metals such as calcium, iron, and zinc leads to an unintended uptake in cells after intake through inhalation and ingestion. In this study we investigated the toxicity and the immunomodulatory potential of Cd in nonactivated and activated murine macrophages (i.e., cell line RAW 264.7). Cadmium alone caused a dose-dependent decreased viability of exposed cells. Subtoxic Cd concentrations delayed cell death in macrophages, resulting from cytotoxic storm, producing reactive oxygen species (ROS) and nitric oxide (NO), in response to their stimulation by bacterial antigens via pattern-recognition receptors (PRRs). In addition, production of selected pro- and anti-inflammatory cytokines, the chemokine CXCL1 (KC), and NO was determined. We observed that proinflammatory IL-1β and also CXCL1 were highly upregulated whereas anti-inflammatory or regulatory cytokines IL-6 and IL-10 were suppressed by 10 µM Cd. Also production of antibacterial NO was significantly reduced through exposure to 10 µM Cd, maybe explaining better survival of macrophages. Additionally, we could show by analysis via ICP-MS that different effects of Cd in nonactivated and activated macrophages definitely did not result from different Cd uptake rates.

No MeSH data available.


Related in: MedlinePlus

Influence of Cd on the cytokine production of RAW 264.7 macrophages. A volume of 10 mL containing 1 × 106 cells/mL was seeded in a culture dish and subsequently stimulated with 0.1 μM or 10 μM Cd in the presence of hk S.E. After indicated periods of incubation supernatants were collected and cytokine (i.e., IL-1β (a), TNF-α (b), IL-6 (c), and IL-10 (d)) concentration was determined by ELISA. Data represent the mean ± S.E.M. and are representative of three independent experiments (n = 3).
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fig3: Influence of Cd on the cytokine production of RAW 264.7 macrophages. A volume of 10 mL containing 1 × 106 cells/mL was seeded in a culture dish and subsequently stimulated with 0.1 μM or 10 μM Cd in the presence of hk S.E. After indicated periods of incubation supernatants were collected and cytokine (i.e., IL-1β (a), TNF-α (b), IL-6 (c), and IL-10 (d)) concentration was determined by ELISA. Data represent the mean ± S.E.M. and are representative of three independent experiments (n = 3).

Mentions: In order to identify immunoregulatory effects of Cd on macrophages the secretion of proinflammatory (i.e., IL-1β and TNF-α) and anti-inflammatory cytokines (IL-10 and IL-6) in response to Cd exposure with or without hk S.E. stimulation were measured by ELISA. Thereby it was of special interest to obtain data that may explain better survival of antigen-stimulated cells in the presence of 10 μM Cd. Therefore, we compared cytokine concentrations in supernatants of cells exposed to 0.1 μM or 10 μM Cd in comparison to untreated controls (Figure 3). The results revealed that in case of IL-1β 0.1 μM Cd exhibited no effect compared to control, whereas 10 μM Cd led to a threefold increase of IL-1β concentration (Figure 3(a)). In terms of IL-10 and IL-6 0.1 μM Cd led to a slightly decreased cytokine secretion, while 10 μM Cd reduced the concentration of both cytokines by half (Figures 3(b) and 3(c)). The TNF-α secretion highly increased after hk S.E. stimulation, whereby the Cd exposition did not result in significant alterations of the cytokine production (Figure 3(d)). The secretion of chemokine CXCL1 was not effected by 0.1 μM Cd compared to control, whereas 10 μM Cd caused twofold increase of CXCL1 concentration in RAW 264.7 culture supernatants (Figure 4(a)). In contrast, NO production was found to be reduced by approximately 40% following exposure to 10 μM Cd; however, 0.1 μM Cd did not show any effect on this functional parameter (Figure 4(b)).


Subtoxic Doses of Cadmium Modulate Inflammatory Properties of Murine RAW 264.7 Macrophages.

Riemschneider S, Herzberg M, Lehmann J - Biomed Res Int (2015)

Influence of Cd on the cytokine production of RAW 264.7 macrophages. A volume of 10 mL containing 1 × 106 cells/mL was seeded in a culture dish and subsequently stimulated with 0.1 μM or 10 μM Cd in the presence of hk S.E. After indicated periods of incubation supernatants were collected and cytokine (i.e., IL-1β (a), TNF-α (b), IL-6 (c), and IL-10 (d)) concentration was determined by ELISA. Data represent the mean ± S.E.M. and are representative of three independent experiments (n = 3).
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig3: Influence of Cd on the cytokine production of RAW 264.7 macrophages. A volume of 10 mL containing 1 × 106 cells/mL was seeded in a culture dish and subsequently stimulated with 0.1 μM or 10 μM Cd in the presence of hk S.E. After indicated periods of incubation supernatants were collected and cytokine (i.e., IL-1β (a), TNF-α (b), IL-6 (c), and IL-10 (d)) concentration was determined by ELISA. Data represent the mean ± S.E.M. and are representative of three independent experiments (n = 3).
Mentions: In order to identify immunoregulatory effects of Cd on macrophages the secretion of proinflammatory (i.e., IL-1β and TNF-α) and anti-inflammatory cytokines (IL-10 and IL-6) in response to Cd exposure with or without hk S.E. stimulation were measured by ELISA. Thereby it was of special interest to obtain data that may explain better survival of antigen-stimulated cells in the presence of 10 μM Cd. Therefore, we compared cytokine concentrations in supernatants of cells exposed to 0.1 μM or 10 μM Cd in comparison to untreated controls (Figure 3). The results revealed that in case of IL-1β 0.1 μM Cd exhibited no effect compared to control, whereas 10 μM Cd led to a threefold increase of IL-1β concentration (Figure 3(a)). In terms of IL-10 and IL-6 0.1 μM Cd led to a slightly decreased cytokine secretion, while 10 μM Cd reduced the concentration of both cytokines by half (Figures 3(b) and 3(c)). The TNF-α secretion highly increased after hk S.E. stimulation, whereby the Cd exposition did not result in significant alterations of the cytokine production (Figure 3(d)). The secretion of chemokine CXCL1 was not effected by 0.1 μM Cd compared to control, whereas 10 μM Cd caused twofold increase of CXCL1 concentration in RAW 264.7 culture supernatants (Figure 4(a)). In contrast, NO production was found to be reduced by approximately 40% following exposure to 10 μM Cd; however, 0.1 μM Cd did not show any effect on this functional parameter (Figure 4(b)).

Bottom Line: Cadmium alone caused a dose-dependent decreased viability of exposed cells.Also production of antibacterial NO was significantly reduced through exposure to 10 µM Cd, maybe explaining better survival of macrophages.Additionally, we could show by analysis via ICP-MS that different effects of Cd in nonactivated and activated macrophages definitely did not result from different Cd uptake rates.

View Article: PubMed Central - PubMed

Affiliation: Fraunhofer Institute for Cell Therapy and Immunology (IZI), 04103 Leipzig, Germany.

ABSTRACT
Cadmium (Cd) is a toxic heavy metal that exhibits various adverse effects in the human and animal organism. Its resemblance to essential metals such as calcium, iron, and zinc leads to an unintended uptake in cells after intake through inhalation and ingestion. In this study we investigated the toxicity and the immunomodulatory potential of Cd in nonactivated and activated murine macrophages (i.e., cell line RAW 264.7). Cadmium alone caused a dose-dependent decreased viability of exposed cells. Subtoxic Cd concentrations delayed cell death in macrophages, resulting from cytotoxic storm, producing reactive oxygen species (ROS) and nitric oxide (NO), in response to their stimulation by bacterial antigens via pattern-recognition receptors (PRRs). In addition, production of selected pro- and anti-inflammatory cytokines, the chemokine CXCL1 (KC), and NO was determined. We observed that proinflammatory IL-1β and also CXCL1 were highly upregulated whereas anti-inflammatory or regulatory cytokines IL-6 and IL-10 were suppressed by 10 µM Cd. Also production of antibacterial NO was significantly reduced through exposure to 10 µM Cd, maybe explaining better survival of macrophages. Additionally, we could show by analysis via ICP-MS that different effects of Cd in nonactivated and activated macrophages definitely did not result from different Cd uptake rates.

No MeSH data available.


Related in: MedlinePlus