TCR-mediated functions are enhanced in activated peripheral blood T cells isolated from leucocyte reduction systems

Tremblay, Mikaela M.; Houtman, Jon C. D.

doi:10.1016/j.jim.2014.11.009

Cited by 7 publications

(5 citation statements)

References 32 publications

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“…Peripheral blood mononuclear cells (PBMCs) were obtained from anonymous donors as previously described [ 17 ]. Blood donors at the DeGowin Blood Center at the University of Iowa Hospitals and Clinics between 18 and 55 years provided written informed consent for cells not used for transfusion to be used for research.…”

Section: Methodsmentioning

confidence: 99%

Exposure of Human CD4 T Cells to IL-12 Results in Enhanced TCR-Induced Cytokine Production, Altered TCR Signaling, and Increased Oxidative Metabolism

et al. 2016

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Human CD4 T cells are constantly exposed to IL-12 during infections and certain autoimmune disorders. The current paradigm is that IL-12 promotes the differentiation of naïve CD4 T cells into Th1 cells, but recent studies suggest IL-12 may play a more complex role in T cell biology. We examined if exposure to IL-12 alters human CD4 T cell responses to subsequent TCR stimulation. We found that IL-12 pretreatment increased TCR-induced IFN-γ, TNF-α, IL-13, IL-4 and IL-10 production. This suggests that prior exposure to IL-12 potentiates the TCR-induced release of a range of cytokines. We observed that IL-12 mediated its effects through both transcriptional and post-transcriptional mechanisms. IL-12 pretreatment increased the phosphorylation of AKT, p38 and LCK following TCR stimulation without altering other TCR signaling molecules, potentially mediating the increase in transcription of cytokines. In addition, the IL-12-mediated enhancement of cytokines that are not transcriptionally regulated was partially driven by increased oxidative metabolism. Our data uncover a novel function of IL-12 in human CD4 T cells; specifically, it enhances the release of a range of cytokines potentially by altering TCR signaling pathways and by enhancing oxidative metabolism.

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Section: Methodsmentioning

confidence: 99%

Exposure of Human CD4 T Cells to IL-12 Results in Enhanced TCR-Induced Cytokine Production, Altered TCR Signaling, and Increased Oxidative Metabolism

et al. 2016

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“…While the blood used in this publication was sourced using typical leukapheresis techniques common for production of blood products, another potential source of raw materials for this system is the leukoreduction cones that are discarded during blood donations to remove host leukocytes which are undesirable for transfusion. There have been multiple publications outlining how these cells could be used to support research and development in cell therapy. − The methods outlined here suggest that these cells could be used to support synthetic biology using a waste product from an existing process. Additionally, the extracts from these donors could be pooled to produce extract that is representative of a population rather than an individual.…”

Section: Discussionmentioning

confidence: 99%

A Cell-Free Protein Expression System Derived from Human Primary Peripheral Blood Mononuclear Cells

et al. 2020

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Historically, some of the first cell-free protein expression systems studied in vitro translation in various human blood cells. However, because of limited knowledge of eukaryotic translation and the advancement of cell line development, interest in these systems decreased. Eukaryotic translation is a complex system of factors that contribute to the overall translation of mRNA to produce proteins. The intracellular translateome of a cell can be modified by various factors and disease states, but it is impossible to individually measure all factors involved when there is no comprehensive understanding of eukaryotic translation. The present work outlines the use of a coupled transcription and translation cell-free protein expression system to produce recombinant proteins derived from human donor peripheral blood mononuclear cells (PBMCs) activated with phytohemagglutinin-M (PHA-M). The methods outlined here could result in tools to aid immunology, gene therapy, cell therapy, and synthetic biology research and provide a convenient and holistic method to study and assess the intracellular translation environment of primary immune cells.

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“…Peripheral blood mononuclear cells (PMBCs) were obtained from anonymous donors as previously described [23]. The CD8 T cells were isolated by negative selection using an enrichment kit for CD8 T cells (Stem Cell Technologies).…”

Section: Methodsmentioning

confidence: 99%

Pretreatment of activated human CD8 T cells with IL-12 leads to enhanced TCR-induced signaling and cytokine production

Vacaflores

Freedman

Chapman

et al. 2017

Molecular Immunology

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During the immune response to pathogens and autoantigens, CD8 T cells are exposed to numerous inflammatory agents including the cytokine IL-12. Previous studies have focused on how IL-12 regulates T cell functions when present during or after the activation of the T cell receptor (TCR). However, recent studies suggest that prior exposure to IL-12 also alters the TCR responsiveness of murine T cells. Whether similar phenomena occur in human activated CD8 T cells and the mechanisms mediating these effects remain unexplored. In this study, we observed that pretreatment of human activated CD8 T cells with IL-12 results in increased cytokine mRNA and protein production following subsequent TCR challenge. The potentiation of TCR-mediated cytokine release was transient and required low doses of IL-12 for at least 24 hours. Mechanistically, prior exposure to IL-12 increased the TCR induced activation of select MAPKs and AKT without altering the activation of more proximal TCR signaling molecules, suggesting that the IL-12 mediated changes in TCR signaling are responsible for the increased production of cytokines. Our data suggest that prior treatment with IL-12 potentiates human CD8 T cell responses at sites of infection and inflammation, expanding our understanding of the function of this clinically important cytokine.

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TCR-mediated functions are enhanced in activated peripheral blood T cells isolated from leucocyte reduction systems

Cited by 7 publications

References 32 publications

Exposure of Human CD4 T Cells to IL-12 Results in Enhanced TCR-Induced Cytokine Production, Altered TCR Signaling, and Increased Oxidative Metabolism

Exposure of Human CD4 T Cells to IL-12 Results in Enhanced TCR-Induced Cytokine Production, Altered TCR Signaling, and Increased Oxidative Metabolism

A Cell-Free Protein Expression System Derived from Human Primary Peripheral Blood Mononuclear Cells

Pretreatment of activated human CD8 T cells with IL-12 leads to enhanced TCR-induced signaling and cytokine production

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