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Expression data from human CD4 or CD8 T-cells isolated from PBMC cultured at a low cell density (LDC) or high cell density (HDC)

ABSTRACT: Research in human immunobiology is mainly based on working with peripheral blood mononuclear cells (PBMC). However, recent investigations have shown that circulating CD4+ T cells are less sensitive to several T-cell activating monoclonal antibodies (mAb) and to recall antigens as compared to tissue-resident cells or cells that were in-vitro cultured at a high cell density of 10^7 cells/mL for 2 days at 37°C and 5% CO2 (RESTORE protocol, Römer et al., Blood 2011, PMID: 21931118). To explain the increase in sensitivity of CD4+ T-cells to mAbs and recall antigens on a molecular level, we performed microarray hybridizations of total RNA from T-cells isolated from PBMC that were cultured at a low or high cell density. To avoid the detection of genes that are up- or down-regulated by the culture process itself, we used low cell density cultured PBMC, instead of freshly prepared PBMC. We used microarrays to detail the global programme of gene expression underlying differences in the cell density of human PBMC and identified genes that are significantly up- or downregulated dependent on the cell density of PBMC.

ORGANISM(S): Homo sapiens

PROVIDER: GSE51288 | GEO | 2015/06/09

SECONDARY ACCESSION(S): PRJNA222314

REPOSITORIES: GEO

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Expression data from human CD4 or CD8 T-cells isolated from PBMC cultured at a low cell density (LDC) or high cell density (HDC)

Project description:Research in human immunobiology is mainly based on working with peripheral blood mononuclear cells (PBMC). However, recent investigations have shown that circulating CD4+ T cells are less sensitive to several T-cell activating monoclonal antibodies (mAb) and to recall antigens as compared to tissue-resident cells or cells that were in-vitro cultured at a high cell density of 10^7 cells/mL for 2 days at 37°C and 5% CO2 (RESTORE protocol, Römer et al., Blood 2011, PMID: 21931118). To explain the increase in sensitivity of CD4+ T-cells to mAbs and recall antigens on a molecular level, we performed microarray hybridizations of total RNA from T-cells isolated from PBMC that were cultured at a low or high cell density. To avoid the detection of genes that are up- or down-regulated by the culture process itself, we used low cell density cultured PBMC, instead of freshly prepared PBMC. We used microarrays to detail the global programme of gene expression underlying differences in the cell density of human PBMC and identified genes that are significantly up- or downregulated dependent on the cell density of PBMC. Human PBMC of one healthy blood donor were cultured at a low cell density (10^6 cells/mL) or at a high cell density (10^7 cells/mL) for 2 days at 37°C and 5% CO2 and CD4 or CD8 T-cells of both cultures were isolated by MACSbeads. Expression profiles from total RNA extracts were generated by hybridization to Affymetrix microarrays.

2015-06-09 | E-GEOD-51288 | biostudies-arrayexpress

Comparison of CD8+ memory T-cells from fresh and high density precultured PBMC.

Project description:High density cultured CD8+ memory T-cells from peripheral blood mononuclear cells (PBMC) were found to be more sensitive towards antigenic stimulation. We used microarrays to detail the global programme of gene expression underlying differences in the cell density of human PBMC and identified genes that are significantly up- or downregulated dependent on the cell density of PBMC.

2015-06-09 | GSE63430 | GEO

Comparison of CD8+ memory T-cells from fresh and high density precultured PBMC.

2015-06-09 | E-GEOD-63430 | biostudies-arrayexpress

CD73+ CD127high long-term memory CD4 T cells are highly proliferative in response to recall antigens and are early targets in HIV-1 infection

Project description:HIV-1 infection rapidly leads to a loss of the proliferative response of memory CD4+ T lymphocytes, when cultured with recall antigens. We report here that CD73 expression defines a subset of resting memory CD4 T cells which highly express the alpha-chain of the receptor for IL-7 (CD127), but not CD38 or Ki-67 in vivo, yet are highly proliferative in response to mitogen and recall antigens, and to IL-7, in vitro. These cells also preferentially express CCR5 and produce IL-2. We reasoned that CD73+ memory CD4+ T cells decrease early in HIV-1 infection. We compared gene expression profiles of healthy subjects recovered CD4+CD73+ to CD4+CD73- T cells, and also CD8+CD73+ to CD8+CD73- T cells

2021-02-02 | GSE160805 | GEO

Pneumococcal conjugate vaccination at birth induces robust recall responses at 9 months of age.

Project description:The objective of this study was to compare recall responses to vaccine antigens at 3 months and 9 months of age in infants who were vaccinated at birth or at 1 month. PBMC were obtained at 9 months of age from infants who were vaccinated at birth (n=25) or 1 month (n=25). The PBMC were cultured in the presence or absence of CRM197 antigen for 72 hours. At the termination of the cultures, total RNA was extracted from the PBMC. The RNA was pooled into groups of n=5 subjects per group, following by labeling and hybridization to Affymetrix microarrays.

2011-06-27 | E-GEOD-25263 | biostudies-arrayexpress

Global gene expression pattern difference between the high density and the regular density cultured rat bone marrow cells

Project description:In vitro expansion of endothelial progenitor cells (EPCs) remains a challenge in stem cell research and its application. We hypothesize that high density culture is able to expansion EPCs from bone marrow by mimicking cell-cell interactions of the bone marrow niche. To test the hypothesis, rat bone marrow cells were either cultured in high density by seeding or cultured in regular density. Flow cytometric analyses of the cells cultured for 15 days showed that high density cells exhibited smaller cell size and higher levels of marker expression related to EPCs when compared to regular density cultured cells. Global gene expression pattern difference between the high density and the regular density cultured cells was analyzed by a microarray assay.

2013-08-03 | GSE49510 | GEO

Global gene expression pattern difference between the high density and the regular density cultured rat bone marrow cells

2013-08-03 | E-GEOD-49510 | biostudies-arrayexpress

Gene expression profiles of human Ad5- and CMV-specific CD4 T cells

Project description:Analysis of global gene expression profiles of FACS-sorted, human Ad5- and CMV-specific CD4 T cells from the same PBMC sample of healthy donros, using affymetrix Human Gene 2.0ST Gene-Chips; The analysis show that Ad5- and CMV-specific CD4 T cells demonstrate differential gene expression profiles even though the cells are from the same PBMC PBMC from healthy donors (n=3) were CFSE-labeled, then equally divided and stimulated with Ad5 and CMV antigens; On day 6 post stimulation, Ag-specific CD4 T cells were sorted by flow based on CFSE-low.

2014-08-07 | E-GEOD-60180 | biostudies-arrayexpress

Homo sapiens

Project description:Expression data from human CD4 or CD8 T-cells isolated from PBMC cultured at a low cell density (LDC) or high cell density (HDC)

| PRJNA222314 | ENA

Genome-wide chromatin accessibility changes in cultured primary human monocytes cultured at low density and high density.

Project description:ATAC sequencing of primary human monocytes cultured at low and high density. Monocytes isolated from PBMCs of 3 healthy donors were cultured at low density (1 x 10^6 cells/mL) or at high density (1 x10^7 cells/mL) for 24hrs and harvested for ATAC sequencing. Protein expression of FcgR2b is higher on monocytes in high density conditions compared to low density conditions, where expression is negligble. This study provides information on genome-wide chromatin accessibility changes that occur in high density culture in order to study associations with FcgR2b expression.

2022-04-13 | GSE166100 | GEO

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