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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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Related in: MedlinePlus

Technology for cell culture experiments in different microgravity research platforms. (a) Fast-rotating two-dimensional (2D) clinostat manufactured by the German Aerospace Center (DLR, Cologne, Germany) was used to provide simulated microgravity. Under the chosen experimental conditions (60 rpm, 4 mm pipette diameter) a maximal residual acceleration of 4 × 10−3 g is achieved at the outer radius of the pipette and decreases towards the center. (b) Experimental hardware structure which consists of an incubator rack to store the cell containers temporarily before the experiment at 37°C (left), an experimental rack in which all active aggregates are accommodated and where the living cells are handled during altered gravity (right) and a cooling rack to temporarily store all cell containers after the injection of the stop/fixation liquid at 4°C until landing (front). (c) Payload of TEXUS-49 sounding rocket tempered and vacuum-resistant container with experiment syringe systems. (d) Plunger unit EUE for SIMBOX (Science in Microgravity Box) incubator system, support structure (housing made of PEEK), which includes three culture chambers and six supply units, two for each culture compartment. Each culture chamber represents an independent loop. The culture chambers filled with medium are closed on the top of the housing by means of polycarbonate specimen window slides, where the adherent cells are attached beforehand. The housing is tightened by silicon sealing and covered by an aluminum plate (cover) fixed with screws.
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fig1: Technology for cell culture experiments in different microgravity research platforms. (a) Fast-rotating two-dimensional (2D) clinostat manufactured by the German Aerospace Center (DLR, Cologne, Germany) was used to provide simulated microgravity. Under the chosen experimental conditions (60 rpm, 4 mm pipette diameter) a maximal residual acceleration of 4 × 10−3 g is achieved at the outer radius of the pipette and decreases towards the center. (b) Experimental hardware structure which consists of an incubator rack to store the cell containers temporarily before the experiment at 37°C (left), an experimental rack in which all active aggregates are accommodated and where the living cells are handled during altered gravity (right) and a cooling rack to temporarily store all cell containers after the injection of the stop/fixation liquid at 4°C until landing (front). (c) Payload of TEXUS-49 sounding rocket tempered and vacuum-resistant container with experiment syringe systems. (d) Plunger unit EUE for SIMBOX (Science in Microgravity Box) incubator system, support structure (housing made of PEEK), which includes three culture chambers and six supply units, two for each culture compartment. Each culture chamber represents an independent loop. The culture chambers filled with medium are closed on the top of the housing by means of polycarbonate specimen window slides, where the adherent cells are attached beforehand. The housing is tightened by silicon sealing and covered by an aluminum plate (cover) fixed with screws.

Mentions: A fast-rotating two-dimensional (2D) clinostat manufactured by the German Aerospace Center (DLR, Cologne, Germany) was used to provide simulated microgravity (Figure 1(a)). The principle of clinorotation-induced microgravity is the rotation of a cell suspension in a serological pipette perpendicular to the Earth's gravity. The microgravity produced is an averaging of the gravity vector, if the clinostat rotates with 40–100 rpm. Under the chosen experimental conditions (60 rpm, 4 mm pipette diameter) a maximal residual acceleration of 4 × 10−3 g is achieved at the outer radius of the serological pipette and decreases towards the center. The clinostat device was placed in an incubator providing constant 37°C. Fifteen serological pipettes rotated at the same time with 60 rpm. 1 g controls were placed at the ground plate of the clinostat without rotation but the same environmental conditions are as the μg samples. The density of the cell suspension was 0.5 × 106/mL (U937 macrophage-like cells), 0.25–0.5 × 106/mL (human primary macrophages), or 0.75 × 106/mL (BV-2 microglial cells) in 1 mL volume each. The duration of the filling procedure was not longer than 10 min for all 30 serological pipettes. Cells were cultured in the serological pipettes for 24 h–96 h. After clinorotation, cells were fixed by the addition of 500 μL of 3% PFA (Sigma-Aldrich)/2% sucrose (Sigma-Aldrich) solution for 30 min, washed with PBS and analyzed after immunocytochemical staining by flow cytometry.


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)

Technology for cell culture experiments in different microgravity research platforms. (a) Fast-rotating two-dimensional (2D) clinostat manufactured by the German Aerospace Center (DLR, Cologne, Germany) was used to provide simulated microgravity. Under the chosen experimental conditions (60 rpm, 4 mm pipette diameter) a maximal residual acceleration of 4 × 10−3 g is achieved at the outer radius of the pipette and decreases towards the center. (b) Experimental hardware structure which consists of an incubator rack to store the cell containers temporarily before the experiment at 37°C (left), an experimental rack in which all active aggregates are accommodated and where the living cells are handled during altered gravity (right) and a cooling rack to temporarily store all cell containers after the injection of the stop/fixation liquid at 4°C until landing (front). (c) Payload of TEXUS-49 sounding rocket tempered and vacuum-resistant container with experiment syringe systems. (d) Plunger unit EUE for SIMBOX (Science in Microgravity Box) incubator system, support structure (housing made of PEEK), which includes three culture chambers and six supply units, two for each culture compartment. Each culture chamber represents an independent loop. The culture chambers filled with medium are closed on the top of the housing by means of polycarbonate specimen window slides, where the adherent cells are attached beforehand. The housing is tightened by silicon sealing and covered by an aluminum plate (cover) fixed with screws.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4309248&req=5

fig1: Technology for cell culture experiments in different microgravity research platforms. (a) Fast-rotating two-dimensional (2D) clinostat manufactured by the German Aerospace Center (DLR, Cologne, Germany) was used to provide simulated microgravity. Under the chosen experimental conditions (60 rpm, 4 mm pipette diameter) a maximal residual acceleration of 4 × 10−3 g is achieved at the outer radius of the pipette and decreases towards the center. (b) Experimental hardware structure which consists of an incubator rack to store the cell containers temporarily before the experiment at 37°C (left), an experimental rack in which all active aggregates are accommodated and where the living cells are handled during altered gravity (right) and a cooling rack to temporarily store all cell containers after the injection of the stop/fixation liquid at 4°C until landing (front). (c) Payload of TEXUS-49 sounding rocket tempered and vacuum-resistant container with experiment syringe systems. (d) Plunger unit EUE for SIMBOX (Science in Microgravity Box) incubator system, support structure (housing made of PEEK), which includes three culture chambers and six supply units, two for each culture compartment. Each culture chamber represents an independent loop. The culture chambers filled with medium are closed on the top of the housing by means of polycarbonate specimen window slides, where the adherent cells are attached beforehand. The housing is tightened by silicon sealing and covered by an aluminum plate (cover) fixed with screws.
Mentions: A fast-rotating two-dimensional (2D) clinostat manufactured by the German Aerospace Center (DLR, Cologne, Germany) was used to provide simulated microgravity (Figure 1(a)). The principle of clinorotation-induced microgravity is the rotation of a cell suspension in a serological pipette perpendicular to the Earth's gravity. The microgravity produced is an averaging of the gravity vector, if the clinostat rotates with 40–100 rpm. Under the chosen experimental conditions (60 rpm, 4 mm pipette diameter) a maximal residual acceleration of 4 × 10−3 g is achieved at the outer radius of the serological pipette and decreases towards the center. The clinostat device was placed in an incubator providing constant 37°C. Fifteen serological pipettes rotated at the same time with 60 rpm. 1 g controls were placed at the ground plate of the clinostat without rotation but the same environmental conditions are as the μg samples. The density of the cell suspension was 0.5 × 106/mL (U937 macrophage-like cells), 0.25–0.5 × 106/mL (human primary macrophages), or 0.75 × 106/mL (BV-2 microglial cells) in 1 mL volume each. The duration of the filling procedure was not longer than 10 min for all 30 serological pipettes. Cells were cultured in the serological pipettes for 24 h–96 h. After clinorotation, cells were fixed by the addition of 500 μL of 3% PFA (Sigma-Aldrich)/2% sucrose (Sigma-Aldrich) solution for 30 min, washed with PBS and analyzed after immunocytochemical staining by flow cytometry.

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