Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of human T-lymphocytes and monocytes from participants of the Tour de France 2005

ABSTRACT: Exercise has an measureable impact on gene expression profiles of white blood cells. While subpopulations of white blood cells undergo specific changes in answer to physical load we focused on pan T-lymphocytes and Monocytes. These cells are in direct contact to organs influenced by exercise and additionally should be affected by alterations of nutrient and gas composition in blood, which occur as a consequence of exercise. We screened the gene expression profiles of top athletes at the beginning and the end of the Tour de France 2005 and at at the annual rest period in order to find functional processes that are differentially expressed in answer to the longtime and high load training and competition. Therefor we filtered out genes with significant changes in gene expression more than 1.5fold between two of the three conditions and build overlap lists of this list and GO (gene ontology) lists and KEGG (kyoto encyclopedia of genes and genomes) lists. We observed a strong decline in immune functions and an equal decline in metabolic processes. Experiment Overall Design: 12 professional cyclists from two german top teams gave written agreement to participate to this study. We collected 12ml of whole blood at each time point and isolated T-lymphocytes out of 4ml and monocytes out of 8ml whole blood. The experimental groups are divided into rest (6 replicates in Tcells/ 7 replicates in Monocytes), start (10 replicates in each cell type) and final (8 replicates in Tcells/ 9 replicates in Monocytes). This means that we collected samples at the beginning and the end of the Tour de France and during the annual rest period of the athletes.

ORGANISM(S): Homo sapiens

SUBMITTER: Sandy Mosig

PROVIDER: E-GEOD-5105 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:White blood cells (WBCs) express tens of thousands of genes. Their expression levels are modified by genetic and external factors. In further study we found that gene expression profiles of these cellls can serve as surrogate markers for monitoring exercise and training load.The purpose of the present study was to investigate the effects of acute exercise on gene expression profiles (GEPs) of PBMCs and T-lymphocytes in order to re-detect the patterns in subpopulations. The use of WBC subpopulations is necessary because of the well known subpopulation shifts that occur during physical activities. Three male probands performed an exhaustive treadmill test (ET) at 80% of their VO2max. PBMCs were isolated using BD Vacutainer CPT tubes containig Ficoll. T-lymphocytes were isolated from the PBMCs via rosetting technology. Gene expression profiles were measured using the Affymetrix GeneChipÂ® technology. After scaling, normalisation, and filtering groupwise and pairwise comparisons of gene expression intensities were performed. We found that sorting increases the detection sensitivity and enables the researcher to observe regulation that is hidden in heterogenous populations. We therefore suggest not to work on mixed nbut instead on sorted cells when performing gene expression analyses. Experiment Overall Design: Three healthy male probands executed an exhaustive treadmill test (80% VO2max) until individual exhaustion. Blood samples (16ml) were drawn before and one hour past the tests. PBMCs were isolated using the Ficoll density centrifugation methode. T-lymphocytes were isolated by adding rosetting anibodies zo the cells before centrifugation. Cells were lysed using Trizol and worked up with Qiagen RNeasy Micro columns. RNA was processed and hybridised on U133A 2.0 Affymetrix GeneChips. Samples were grouped and gene expression changes were detected via multiple algorithms included in GeneSpring 7.2 (Agilent). Experiment Overall Design: The four groups contained three samples related to the cell type "PBMC" or "T-cell" and "before exercise" or "1hour past exercise". TTest, GOs and KEGG classification, principal component analysis and hierarchical clstering were used to scan for gene expression profiles induced through the different exercise conditions.

Project description:White blood cells (WBCs) express tens of thousands of genes. Their expression levels are modified by genetic and external factors. The purpose of the present study was to investigate the effects of acute exercise on gene expression profiles (GEPs) of WBCs and to identify suitable genes which may serve as surrogate markers for monitoring exercise and training load. Five male probands performed an exhaustive treadmill test (ET) at 80% of their VO2max, and a moderate treadmill test (MT) at 60% VO2max for exactly the same time one to two week later. White blood cells (WBCs) were isolated by the erythrocyte lysis method. Gene expression profiles were measured using the Affymetrix GeneChipÂ® technology. After scaling, normalisation, and filtering groupwise and pairwise comparisons of gene expression intensities were performed and validated by real-time PCR. We found 450 genes up-regulated and 150 down-regulated (> 1.5-fold change; ANOVA with Benjamini-Hochberg correction for multiple testing, p<0.05) upon exhaustive exercise. Analysis of mean expression levels after MT showed that the extent of up- and down-regulation was workload dependent. The genes for the stress proteins HSPA1A, HSPH1 and the matrix metalloproteinase MMP-9 showed the most prominent increases whereas the mRNA concentrations of the YES1 oncogene (YES1), of CD160 (BY55), and of a member of the mitochondrial electron transport chain (ATP5s) were most strongly reduced. Remarkably, we could largely reproduce the data from a previous report by Connolly et al. (01) even though we used considerably different methodology. After acute exercise the genes with increased expression levels were highly significantly associated with the gene ontology terms heat-shock proteins, apoptosis and inflammation. The results demonstrate that gene expression changes in WBCs can reflect intensity and duration of exercise. Further analysis is needed to confirm the applicability of expression fingerprints as useful tools for monitoring exercise and training loads in order to avoid training associated health risks. Experiment Overall Design: Five healthy male probands executed an exhaustive treadmill test (80% VO2max) until individual exhaustion. One week later they repeated the test at 60% VO2max for the same time (moderate test). Blood samples (9ml) were drawn before and one hour past the tests. Erythrocytes were lysed and RNA was isolated from the white blood cells. RNA was processed and hybridised on U133A 2.0 Affyetrix GeneChips. Samples were grouped and gene expression changes were detected via multiple algorithms included in GeneSpring 7.2 (Agilent). Experiment Overall Design: Three groups were build from the twenty samples. All ten pre exercise samples were grouped together as the pre test" group. The other two groups contained five samples related to the "exhaustive test" or the "moderate test". TTest in cobination with multiple testing corrections, principal component analysis and hierarchical clstering were used to scan for gene expression profiles induced through the different exercise conditions.

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Transcription profiling of human T-lymphocytes and monocytes from participants of the Tour de France 2005

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