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Rapid alterations of cell cycle control proteins in human T lymphocytes in microgravity.

Thiel CS, Paulsen K, Bradacs G, Lust K, Tauber S, Dumrese C, Hilliger A, Schoppmann K, Biskup J, Gölz N, Sang C, Ziegler U, Grote KH, Zipp F, Zhuang F, Engelmann F, Hemmersbach R, Cogoli A, Ullrich O - Cell Commun. Signal (2012)

Bottom Line: In our study we aimed to identify rapidly reacting gravity-responsive mechanisms in mammalian cells in order to understand if and how altered gravity is translated into a cellular response.Additionally, during 2D clinorotation, Tyr-15-phosphorylation occurred later and was shorter than in the 1 g controls.The histone acetyltransferase (HAT) inhibitor curcumin was able to abrogate microgravity-induced p21 mRNA expression, whereas expression was enhanced by a histone deacetylase (HDAC) inhibitor.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Anatomy, Faculty of Medicine, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland. oliver.ullrich@anatom.uzh.ch.

ABSTRACT
In our study we aimed to identify rapidly reacting gravity-responsive mechanisms in mammalian cells in order to understand if and how altered gravity is translated into a cellular response. In a combination of experiments using "functional weightlessness" provided by 2D-clinostats and real microgravity provided by several parabolic flight campaigns and compared to in-flight-1g-controls, we identified rapid gravity-responsive reactions inside the cell cycle regulatory machinery of human T lymphocytes. In response to 2D clinorotation, we detected an enhanced expression of p21 Waf1/Cip1 protein within minutes, less cdc25C protein expression and enhanced Ser147-phosphorylation of cyclinB1 after CD3/CD28 stimulation. Additionally, during 2D clinorotation, Tyr-15-phosphorylation occurred later and was shorter than in the 1 g controls. In CD3/CD28-stimulated primary human T cells, mRNA expression of the cell cycle arrest protein p21 increased 4.1-fold after 20s real microgravity in primary CD4+ T cells and 2.9-fold in Jurkat T cells, compared to 1 g in-flight controls after CD3/CD28 stimulation. The histone acetyltransferase (HAT) inhibitor curcumin was able to abrogate microgravity-induced p21 mRNA expression, whereas expression was enhanced by a histone deacetylase (HDAC) inhibitor. Therefore, we suppose that cell cycle progression in human T lymphocytes requires Earth gravity and that the disturbed expression of cell cycle regulatory proteins could contribute to the breakdown of the human immune system in space.

No MeSH data available.


Related in: MedlinePlus

Protein expression and phosphorylation of cdc2, cdc25 and cyclinB1 in Jurkat T cells at the onset (0 min) and after 1 min, 5 min and 10 min simulated weightlessness (μg) without (con) or with PMA (PMA) or CD3/CD28 soluble antibodies (CD3/CD28). 1g-control experiments have been performed inside the clinostat, but without rotation. Three independent experiments were performed and analysed separately by immunoblotting. Quantitation was performed by Gene Profiler software. The results are expressed as mean ± SEM. Data were analysed by one way ANOVA, followed by Bonferroni test for comparison of certain column pairs. p < 0.1 = *, p < 0.01 = **, p < 0.001 = ***
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Figure 3: Protein expression and phosphorylation of cdc2, cdc25 and cyclinB1 in Jurkat T cells at the onset (0 min) and after 1 min, 5 min and 10 min simulated weightlessness (μg) without (con) or with PMA (PMA) or CD3/CD28 soluble antibodies (CD3/CD28). 1g-control experiments have been performed inside the clinostat, but without rotation. Three independent experiments were performed and analysed separately by immunoblotting. Quantitation was performed by Gene Profiler software. The results are expressed as mean ± SEM. Data were analysed by one way ANOVA, followed by Bonferroni test for comparison of certain column pairs. p < 0.1 = *, p < 0.01 = **, p < 0.001 = ***

Mentions: Because long-term in vitro studies clearly revealed that T cells lost their proliferative capacity in microgravity [8,9], we first investigated key molecules of cell cycle control in short-term simulated weightlessness provided by 2D clinorotation of PMA-activated or non-activated human Jurkat T lymphocytes (Figure 1, 2, 3). The first set of experiments aimed to give a first impression on possible rapid and early alterations in the cell cycle control machinery in T cells. Due to the construction principle of the DLR-clinostat, reliable incubation times are minutes long and therefore, technically we were not able to perform clinorotation experiments with shorter incubation times than 1 min.


Rapid alterations of cell cycle control proteins in human T lymphocytes in microgravity.

Thiel CS, Paulsen K, Bradacs G, Lust K, Tauber S, Dumrese C, Hilliger A, Schoppmann K, Biskup J, Gölz N, Sang C, Ziegler U, Grote KH, Zipp F, Zhuang F, Engelmann F, Hemmersbach R, Cogoli A, Ullrich O - Cell Commun. Signal (2012)

Protein expression and phosphorylation of cdc2, cdc25 and cyclinB1 in Jurkat T cells at the onset (0 min) and after 1 min, 5 min and 10 min simulated weightlessness (μg) without (con) or with PMA (PMA) or CD3/CD28 soluble antibodies (CD3/CD28). 1g-control experiments have been performed inside the clinostat, but without rotation. Three independent experiments were performed and analysed separately by immunoblotting. Quantitation was performed by Gene Profiler software. The results are expressed as mean ± SEM. Data were analysed by one way ANOVA, followed by Bonferroni test for comparison of certain column pairs. p < 0.1 = *, p < 0.01 = **, p < 0.001 = ***
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3275513&req=5

Figure 3: Protein expression and phosphorylation of cdc2, cdc25 and cyclinB1 in Jurkat T cells at the onset (0 min) and after 1 min, 5 min and 10 min simulated weightlessness (μg) without (con) or with PMA (PMA) or CD3/CD28 soluble antibodies (CD3/CD28). 1g-control experiments have been performed inside the clinostat, but without rotation. Three independent experiments were performed and analysed separately by immunoblotting. Quantitation was performed by Gene Profiler software. The results are expressed as mean ± SEM. Data were analysed by one way ANOVA, followed by Bonferroni test for comparison of certain column pairs. p < 0.1 = *, p < 0.01 = **, p < 0.001 = ***
Mentions: Because long-term in vitro studies clearly revealed that T cells lost their proliferative capacity in microgravity [8,9], we first investigated key molecules of cell cycle control in short-term simulated weightlessness provided by 2D clinorotation of PMA-activated or non-activated human Jurkat T lymphocytes (Figure 1, 2, 3). The first set of experiments aimed to give a first impression on possible rapid and early alterations in the cell cycle control machinery in T cells. Due to the construction principle of the DLR-clinostat, reliable incubation times are minutes long and therefore, technically we were not able to perform clinorotation experiments with shorter incubation times than 1 min.

Bottom Line: In our study we aimed to identify rapidly reacting gravity-responsive mechanisms in mammalian cells in order to understand if and how altered gravity is translated into a cellular response.Additionally, during 2D clinorotation, Tyr-15-phosphorylation occurred later and was shorter than in the 1 g controls.The histone acetyltransferase (HAT) inhibitor curcumin was able to abrogate microgravity-induced p21 mRNA expression, whereas expression was enhanced by a histone deacetylase (HDAC) inhibitor.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Anatomy, Faculty of Medicine, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland. oliver.ullrich@anatom.uzh.ch.

ABSTRACT
In our study we aimed to identify rapidly reacting gravity-responsive mechanisms in mammalian cells in order to understand if and how altered gravity is translated into a cellular response. In a combination of experiments using "functional weightlessness" provided by 2D-clinostats and real microgravity provided by several parabolic flight campaigns and compared to in-flight-1g-controls, we identified rapid gravity-responsive reactions inside the cell cycle regulatory machinery of human T lymphocytes. In response to 2D clinorotation, we detected an enhanced expression of p21 Waf1/Cip1 protein within minutes, less cdc25C protein expression and enhanced Ser147-phosphorylation of cyclinB1 after CD3/CD28 stimulation. Additionally, during 2D clinorotation, Tyr-15-phosphorylation occurred later and was shorter than in the 1 g controls. In CD3/CD28-stimulated primary human T cells, mRNA expression of the cell cycle arrest protein p21 increased 4.1-fold after 20s real microgravity in primary CD4+ T cells and 2.9-fold in Jurkat T cells, compared to 1 g in-flight controls after CD3/CD28 stimulation. The histone acetyltransferase (HAT) inhibitor curcumin was able to abrogate microgravity-induced p21 mRNA expression, whereas expression was enhanced by a histone deacetylase (HDAC) inhibitor. Therefore, we suppose that cell cycle progression in human T lymphocytes requires Earth gravity and that the disturbed expression of cell cycle regulatory proteins could contribute to the breakdown of the human immune system in space.

No MeSH data available.


Related in: MedlinePlus