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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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ICAM-1 expression in macrophage-like differentiated U937 cells after long-term exposure to microgravity during the SIMBOX/Shenzhou-8 mission. Cells were cultured under standard cell culture conditions (incubator control) or exposed to different gravity conditions during the SIMBOX/Shenzhou-8 mission. Differentiated U937 cells were fixed in microgravity (μg group) or in 1 g (1 g control group) after 5 days. Only CellMask-positive and TUNEL-negative cells were analyzed. (a) Each group represents analysis of the mean fluorescence of 200–1000 individual cells from one recovered slide. Data are expressed as the median of mean single cell fluorescence intensities with the smallest observation (sample minimum), lower quartile, median, upper quartile, and largest observation (sample maximum). Statistical analysis was performed with GraphPad Prism 5, Wilcoxon test, *P < 0.05, **P < 0.01, and ***P < 0.001. (b) Standard cell culture control (con), 1 g hardware control (1 g) and the microgravity sample (μg).
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fig6: ICAM-1 expression in macrophage-like differentiated U937 cells after long-term exposure to microgravity during the SIMBOX/Shenzhou-8 mission. Cells were cultured under standard cell culture conditions (incubator control) or exposed to different gravity conditions during the SIMBOX/Shenzhou-8 mission. Differentiated U937 cells were fixed in microgravity (μg group) or in 1 g (1 g control group) after 5 days. Only CellMask-positive and TUNEL-negative cells were analyzed. (a) Each group represents analysis of the mean fluorescence of 200–1000 individual cells from one recovered slide. Data are expressed as the median of mean single cell fluorescence intensities with the smallest observation (sample minimum), lower quartile, median, upper quartile, and largest observation (sample maximum). Statistical analysis was performed with GraphPad Prism 5, Wilcoxon test, *P < 0.05, **P < 0.01, and ***P < 0.001. (b) Standard cell culture control (con), 1 g hardware control (1 g) and the microgravity sample (μg).

Mentions: During the SIMBOX (Science in Microgravity Box) mission on Shenzhou-8, we investigated microgravity-associated long-term alterations in macrophage-like differentiated U937 cells and analyzed the effect of long-term microgravity on the cytoskeleton and immunologically relevant surface molecules [23]. Human U937 cells were differentiated into a macrophage-like phenotype and exposed to microgravity or 1 g on a reference centrifuge on orbit for 5 days. The unmanned Shenzhou-8 spacecraft was launched with a Long March 2F (CZ-2F) rocket from the Jiuquan Satellite Launch Center (JSLC) and landed after a 17-day mission. After on-orbit fixation, the samples were analyzed with immunocytochemical staining and confocal microscopy after landing. Double fluorescent staining was performed using HCS CellMask deep red for the delineation of cells and FITC-labeled anti ICAM-1 antibody for identification of cell surface expression of ICAM-1. Cells were analyzed as described above. We detected a significant higher expression of ICAM-1 in long-term microgravity in comparison to the in-flight 1 g control group (Figure 6). Similar to the parabolic flight experiments, incubation of the macrophage-like differentiated U937 cells in the experiment hardware caused a significant downregulation of ICAM-1 expression. Thus, it can be excluded that the microgravity effects on ICAM-1 were caused by the experiment system itself.


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)

ICAM-1 expression in macrophage-like differentiated U937 cells after long-term exposure to microgravity during the SIMBOX/Shenzhou-8 mission. Cells were cultured under standard cell culture conditions (incubator control) or exposed to different gravity conditions during the SIMBOX/Shenzhou-8 mission. Differentiated U937 cells were fixed in microgravity (μg group) or in 1 g (1 g control group) after 5 days. Only CellMask-positive and TUNEL-negative cells were analyzed. (a) Each group represents analysis of the mean fluorescence of 200–1000 individual cells from one recovered slide. Data are expressed as the median of mean single cell fluorescence intensities with the smallest observation (sample minimum), lower quartile, median, upper quartile, and largest observation (sample maximum). Statistical analysis was performed with GraphPad Prism 5, Wilcoxon test, *P < 0.05, **P < 0.01, and ***P < 0.001. (b) Standard cell culture control (con), 1 g hardware control (1 g) and the microgravity sample (μg).
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig6: ICAM-1 expression in macrophage-like differentiated U937 cells after long-term exposure to microgravity during the SIMBOX/Shenzhou-8 mission. Cells were cultured under standard cell culture conditions (incubator control) or exposed to different gravity conditions during the SIMBOX/Shenzhou-8 mission. Differentiated U937 cells were fixed in microgravity (μg group) or in 1 g (1 g control group) after 5 days. Only CellMask-positive and TUNEL-negative cells were analyzed. (a) Each group represents analysis of the mean fluorescence of 200–1000 individual cells from one recovered slide. Data are expressed as the median of mean single cell fluorescence intensities with the smallest observation (sample minimum), lower quartile, median, upper quartile, and largest observation (sample maximum). Statistical analysis was performed with GraphPad Prism 5, Wilcoxon test, *P < 0.05, **P < 0.01, and ***P < 0.001. (b) Standard cell culture control (con), 1 g hardware control (1 g) and the microgravity sample (μg).
Mentions: During the SIMBOX (Science in Microgravity Box) mission on Shenzhou-8, we investigated microgravity-associated long-term alterations in macrophage-like differentiated U937 cells and analyzed the effect of long-term microgravity on the cytoskeleton and immunologically relevant surface molecules [23]. Human U937 cells were differentiated into a macrophage-like phenotype and exposed to microgravity or 1 g on a reference centrifuge on orbit for 5 days. The unmanned Shenzhou-8 spacecraft was launched with a Long March 2F (CZ-2F) rocket from the Jiuquan Satellite Launch Center (JSLC) and landed after a 17-day mission. After on-orbit fixation, the samples were analyzed with immunocytochemical staining and confocal microscopy after landing. Double fluorescent staining was performed using HCS CellMask deep red for the delineation of cells and FITC-labeled anti ICAM-1 antibody for identification of cell surface expression of ICAM-1. Cells were analyzed as described above. We detected a significant higher expression of ICAM-1 in long-term microgravity in comparison to the in-flight 1 g control group (Figure 6). Similar to the parabolic flight experiments, incubation of the macrophage-like differentiated U937 cells in the experiment hardware caused a significant downregulation of ICAM-1 expression. Thus, it can be excluded that the microgravity effects on ICAM-1 were caused by the experiment system itself.

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