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Comparison of the effects of GMCSF, IL-4 and 15kDa Granulysin on elutriated monocytes: a 48 hour time point analysis

ABSTRACT: Gene expression profiling of elutriated monocytes cultured for 48 hours in RPMI-1640, 10% AB plamsa and one of the following: GMCSF (56IU/ml), IL-4 (2000 IU/ml), 15 kDa Granulysin (10 nM) Sources tested: monocytes form 3 donors tested at time 0, 4 hours, 12 hours, 24 hours, 48 hours 5 time points were compared: day 0 (cells before start the culture), 4, 12, 24, 48 hours

ORGANISM(S): Homo sapiens

SUBMITTER: Luciano Castiello

PROVIDER: E-GEOD-35304 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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15 kDa granulysin causes differentiation of monocytes to dendritic cells but lacks cytotoxic activity.

Clayberger Carol C Finn Michael W MW Wang Tianhong T Saini Reena R Saini Reena R Wilson Christine C Barr Valarie A VA Sabatino Marianna M Castiello Luciano L Stroncek David D Krensky Alan M AM

Journal of immunology (Baltimore, Md. : 1950) 20120514 12

Granulysin is expressed as two isoforms by human cytotoxic cells: a single mRNA gives rise to 15 kDa granulysin, a portion of which is cleaved to a 9 kDa protein. Studies with recombinant 9 kDa granulysin have demonstrated its cytolytic and proinflammatory properties, but much less is known about the biologic function of the 15 kDa isoform. In this study, we show that the subcellular localization and functions of 9 and 15 kDa granulysin are largely distinct. Nine kilodalton granulysin is confine ...[more]

PMID: 22586033

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Comparison of the effects of GMCSF, IL-4 and 15kDa Granulysin on elutriated monocytes: a 48 hour time point analysis

Project description:Gene expression profiling of elutriated monocytes cultured for 48 hours in RPMI-1640, 10% AB plamsa and one of the following: GMCSF (56IU/ml), IL-4 (2000 IU/ml), 15 kDa Granulysin (10 nM) Sources tested: monocytes form 3 donors tested at time 0, 4 hours, 12 hours, 24 hours, 48 hours

2012-05-19 | GSE35304 | GEO

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Project description:Expression profiles at different time points during dendritic cell differentiation (induced by specific culture conditions) including monocytes as well as expression profiles between monocytes and completely differentiated cells (macrophages at day7 and dendritic cells at day7, respectively) were compared. Monocyte-derived dendritic cells (DC) were obtained by culturing elutriated monocytes with 20U/ml IL-4, 280U/ml GM-CSF and 10% FCS; monocyte-derived macrophages (MAC) were obtained by culturing elutriated monocytes with 2% AB serum. Three to seven biological replicates that are derived from independent healthy donors were included. One-color based gene expression. 2 datasets: dendritic cell kinetic study and comparison of monocyte, macrophage, and dendritic cells

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Homo sapiens

Project description:Comparison of the effects of GMCSF, IL-4 and 15kDa Granulysin on elutriated monocytes: a 48 hour time point analysis

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Project description:A growing body of evidence suggests that inflammatory cytokines have a dualistic role in immunity. In this study, we sought to determine the direct effects IFN-gamma on the differentiation and maturation of human peripheral blood monocyte-derived dendritic cells (moDC). Here, we report that following differentiation of human peripheral-blood monocytes into moDCs with granulocyte-macrophage colony-stimulating factor (GM-CSF) and IL-4, interferon-gamma (IFN-gamma) induces moDC maturation and up-regulates the co-stimulatory markers CD80, CD86, CD95, and MHC Class I, enabling moDCs to effectively generate antigen-specific CD4+ and CD8+ T cell responses for multiple viral and tumor antigens. Interestingly, early exposure of monocytes to high concentrations of IFN-gamma promotes monocyte differentiation into macrophages, despite the presence of GM-CSF and IL-4. However, under low concentrations of IFN-gamma, monocytes continue to differentiate into dendritic cells possessing a unique gene-expression profile, resulting in impairments in subsequent maturation by IFN-gamma and an inability to generate effective antigen-specific CD4+ and CD8+ T cell responses compared to standard moDCs. Monocytes differentiated in the presence of low levels of IFN-gamma downregulate IFN-gamma receptor expression, impairing their response to an inflammatory rechallenge. These findings demonstrate the ability of IFN-gamma to impart differential programs on human moDCs which shape the antigen-specific T cell responses they induce. Timing and intensity of exposure to IFN-gamma can thus determine whether moDCs are tolerogenic or immunostimulating. Human monocyte-derived dendritic cells from 4 healthy donors were differentiated with either GM-CSF and IL-4 (n=4) or GM-CSF, IL-4, and IFN-gamma (n=4). These samples were subsequently hybridized to arrays as 4 biological repeats for each of the two treatment conditions.

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