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Regulation of ICAM-1 in cells of the monocyte/macrophage system in microgravity.

Paulsen K, Tauber S, Dumrese C, Bradacs G, Simmet DM, Gölz N, Hauschild S, Raig C, Engeli S, Gutewort A, Hürlimann E, Biskup J, Unverdorben F, Rieder G, Hofmänner D, Mutschler L, Krammer S, Buttron I, Philpot C, Huge A, Lier H, Barz I, Engelmann F, Layer LE, Thiel CS, Ullrich O - Biomed Res Int (2015)

Bottom Line: In contrast, ICAM-1 expression increased in macrophage-like differentiated human U937 cells during the microgravity phase of parabolic flights and in long-term microgravity provided by a 2D clinostat or during the orbital SIMBOX/Shenzhou-8 mission.In nondifferentiated U937 cells, no effect of microgravity on ICAM-1 expression could be observed during parabolic flight experiments.Thus, ICAM-1 can be considered as a rapid-reacting and sustained gravity-regulated molecule in mammalian cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Anatomy, Faculty of Medicine, University of Zurich, Winterthurerstraß 190, 8057 Zurich, Switzerland.

ABSTRACT
Cells of the immune system are highly sensitive to altered gravity, and the monocyte as well as the macrophage function is proven to be impaired under microgravity conditions. In our study, we investigated the surface expression of ICAM-1 protein and expression of ICAM-1 mRNA in cells of the monocyte/macrophage system in microgravity during clinostat, parabolic flight, sounding rocket, and orbital experiments. In murine BV-2 microglial cells, we detected a downregulation of ICAM-1 expression in clinorotation experiments and a rapid and reversible downregulation in the microgravity phase of parabolic flight experiments. In contrast, ICAM-1 expression increased in macrophage-like differentiated human U937 cells during the microgravity phase of parabolic flights and in long-term microgravity provided by a 2D clinostat or during the orbital SIMBOX/Shenzhou-8 mission. In nondifferentiated U937 cells, no effect of microgravity on ICAM-1 expression could be observed during parabolic flight experiments. We conclude that disturbed immune function in microgravity could be a consequence of ICAM-1 modulation in the monocyte/macrophage system, which in turn could have a strong impact on the interaction with T lymphocytes and cell migration. Thus, ICAM-1 can be considered as a rapid-reacting and sustained gravity-regulated molecule in mammalian cells.

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Cytometry analysis of ICAM-1 expression in macrophage-like differentiated U937 cells or primary human macrophages in simulated microgravity (2D clinorotation). Macrophage-like differentiated U937 cells (a) or primary human macrophages (b) were exposed to either clinorotation (μg), placed in the clinostat but not rotated (1 g control group), or cultured under standard cell culture conditions (incubator control). Cells were stained for ICAM-1 surface expression and analyzed by flow cytometry. The level of ICAM-1 surface expression is represented by the mean fluorescent intensity assessed by flow cytometry. (a) Quantification of ICAM-1 expression in macrophage-like differentiated U937 cells after exposure to different gravity conditions for 1 h, 3 h, or 5 h, n = 6. (b) Quantification of ICAM-1 expression in primary human macrophages after exposure to different gravity conditions for 1 h, 3 h, or 5 h, n = 3. Data are given as median ± SE (*P < 0.1, **P < 0.05, ***P < 0.01, according to one-way ANOVA followed by Wilcoxon or unpaired t-test).
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fig4: Cytometry analysis of ICAM-1 expression in macrophage-like differentiated U937 cells or primary human macrophages in simulated microgravity (2D clinorotation). Macrophage-like differentiated U937 cells (a) or primary human macrophages (b) were exposed to either clinorotation (μg), placed in the clinostat but not rotated (1 g control group), or cultured under standard cell culture conditions (incubator control). Cells were stained for ICAM-1 surface expression and analyzed by flow cytometry. The level of ICAM-1 surface expression is represented by the mean fluorescent intensity assessed by flow cytometry. (a) Quantification of ICAM-1 expression in macrophage-like differentiated U937 cells after exposure to different gravity conditions for 1 h, 3 h, or 5 h, n = 6. (b) Quantification of ICAM-1 expression in primary human macrophages after exposure to different gravity conditions for 1 h, 3 h, or 5 h, n = 3. Data are given as median ± SE (*P < 0.1, **P < 0.05, ***P < 0.01, according to one-way ANOVA followed by Wilcoxon or unpaired t-test).

Mentions: To corroborate the relevance of the results obtained with murine BV-2 microglial cells, we investigated a human macrophage-like cell system. Therefore, human monocytic U937 cells were differentiated into macrophage-like cells [23] and human M2 macrophages were differentiated from blood mononuclear cells. Before the experiment, differentiated macrophage-like cells were detached, resuspended in fresh medium, and filled into 1 mL standardized serological pipettes for the clinostat. Clinorotation was performed for 1 d, 3 d, and 5 d. The 1 g control group of differentiated U937 cells was filled into 1 mL serological pipettes in the same way as the clinorotation cell group but was not rotated. Cells were subsequently fixed and stained for cell surface ICAM-1 and apoptosis (TUNEL) to exclude apoptotic cells from the analysis and subjected to flow cytometry. We detected a highly significant increase of ICAM-1 expression in the clinorotated cells (μg group) compared to the nonrotated cells (1 g control group) after 1 d and 5 d in differentiated U937 cells (Figure 4(a)) and primary macrophages (Figure 4(b)). However, this increase receded after 3 and 5 days of clinorotation. Therefore, we conclude that ICAM-1 expression is increased in human macrophages after 1 and 5 days of simulated microgravity.


Regulation of ICAM-1 in cells of the monocyte/macrophage system in microgravity.

Paulsen K, Tauber S, Dumrese C, Bradacs G, Simmet DM, Gölz N, Hauschild S, Raig C, Engeli S, Gutewort A, Hürlimann E, Biskup J, Unverdorben F, Rieder G, Hofmänner D, Mutschler L, Krammer S, Buttron I, Philpot C, Huge A, Lier H, Barz I, Engelmann F, Layer LE, Thiel CS, Ullrich O - Biomed Res Int (2015)

Cytometry analysis of ICAM-1 expression in macrophage-like differentiated U937 cells or primary human macrophages in simulated microgravity (2D clinorotation). Macrophage-like differentiated U937 cells (a) or primary human macrophages (b) were exposed to either clinorotation (μg), placed in the clinostat but not rotated (1 g control group), or cultured under standard cell culture conditions (incubator control). Cells were stained for ICAM-1 surface expression and analyzed by flow cytometry. The level of ICAM-1 surface expression is represented by the mean fluorescent intensity assessed by flow cytometry. (a) Quantification of ICAM-1 expression in macrophage-like differentiated U937 cells after exposure to different gravity conditions for 1 h, 3 h, or 5 h, n = 6. (b) Quantification of ICAM-1 expression in primary human macrophages after exposure to different gravity conditions for 1 h, 3 h, or 5 h, n = 3. Data are given as median ± SE (*P < 0.1, **P < 0.05, ***P < 0.01, according to one-way ANOVA followed by Wilcoxon or unpaired t-test).
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig4: Cytometry analysis of ICAM-1 expression in macrophage-like differentiated U937 cells or primary human macrophages in simulated microgravity (2D clinorotation). Macrophage-like differentiated U937 cells (a) or primary human macrophages (b) were exposed to either clinorotation (μg), placed in the clinostat but not rotated (1 g control group), or cultured under standard cell culture conditions (incubator control). Cells were stained for ICAM-1 surface expression and analyzed by flow cytometry. The level of ICAM-1 surface expression is represented by the mean fluorescent intensity assessed by flow cytometry. (a) Quantification of ICAM-1 expression in macrophage-like differentiated U937 cells after exposure to different gravity conditions for 1 h, 3 h, or 5 h, n = 6. (b) Quantification of ICAM-1 expression in primary human macrophages after exposure to different gravity conditions for 1 h, 3 h, or 5 h, n = 3. Data are given as median ± SE (*P < 0.1, **P < 0.05, ***P < 0.01, according to one-way ANOVA followed by Wilcoxon or unpaired t-test).
Mentions: To corroborate the relevance of the results obtained with murine BV-2 microglial cells, we investigated a human macrophage-like cell system. Therefore, human monocytic U937 cells were differentiated into macrophage-like cells [23] and human M2 macrophages were differentiated from blood mononuclear cells. Before the experiment, differentiated macrophage-like cells were detached, resuspended in fresh medium, and filled into 1 mL standardized serological pipettes for the clinostat. Clinorotation was performed for 1 d, 3 d, and 5 d. The 1 g control group of differentiated U937 cells was filled into 1 mL serological pipettes in the same way as the clinorotation cell group but was not rotated. Cells were subsequently fixed and stained for cell surface ICAM-1 and apoptosis (TUNEL) to exclude apoptotic cells from the analysis and subjected to flow cytometry. We detected a highly significant increase of ICAM-1 expression in the clinorotated cells (μg group) compared to the nonrotated cells (1 g control group) after 1 d and 5 d in differentiated U937 cells (Figure 4(a)) and primary macrophages (Figure 4(b)). However, this increase receded after 3 and 5 days of clinorotation. Therefore, we conclude that ICAM-1 expression is increased in human macrophages after 1 and 5 days of simulated microgravity.

Bottom Line: In contrast, ICAM-1 expression increased in macrophage-like differentiated human U937 cells during the microgravity phase of parabolic flights and in long-term microgravity provided by a 2D clinostat or during the orbital SIMBOX/Shenzhou-8 mission.In nondifferentiated U937 cells, no effect of microgravity on ICAM-1 expression could be observed during parabolic flight experiments.Thus, ICAM-1 can be considered as a rapid-reacting and sustained gravity-regulated molecule in mammalian cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Anatomy, Faculty of Medicine, University of Zurich, Winterthurerstraß 190, 8057 Zurich, Switzerland.

ABSTRACT
Cells of the immune system are highly sensitive to altered gravity, and the monocyte as well as the macrophage function is proven to be impaired under microgravity conditions. In our study, we investigated the surface expression of ICAM-1 protein and expression of ICAM-1 mRNA in cells of the monocyte/macrophage system in microgravity during clinostat, parabolic flight, sounding rocket, and orbital experiments. In murine BV-2 microglial cells, we detected a downregulation of ICAM-1 expression in clinorotation experiments and a rapid and reversible downregulation in the microgravity phase of parabolic flight experiments. In contrast, ICAM-1 expression increased in macrophage-like differentiated human U937 cells during the microgravity phase of parabolic flights and in long-term microgravity provided by a 2D clinostat or during the orbital SIMBOX/Shenzhou-8 mission. In nondifferentiated U937 cells, no effect of microgravity on ICAM-1 expression could be observed during parabolic flight experiments. We conclude that disturbed immune function in microgravity could be a consequence of ICAM-1 modulation in the monocyte/macrophage system, which in turn could have a strong impact on the interaction with T lymphocytes and cell migration. Thus, ICAM-1 can be considered as a rapid-reacting and sustained gravity-regulated molecule in mammalian cells.

Show MeSH
Related in: MedlinePlus