Cytokine Gene Expression in Normal Human Lymphocytes in Response to Stimulation

Abstract

Sequential gene expression of two type 1 cytokines (interleukin 2 [IL-2] and gamma interferon), one type 2 cytokine (IL-10), two monokines (IL-6 and tumor necrosis factor alpha), and one cytokine receptor (IL-2 receptor [IL-2R]) in normal human peripheral blood mononuclear cells (PBMC) following in vitro stimulation was investigated by reverse transcription-PCR methods. Two stimuli were utilized: phytohemagglutinin (PHA), which acts on the CD2 molecule and T-cell receptors, and anti-CD3 monoclonal antibody, which acts on the CD3 molecule and on T-cell receptors. Increased expression of all studied genes occurred between 1 and 4 hours after stimulation, except for that of the gene encoding IL-10, which was delayed. Expression of all but one of the genes was transient, with a maximal mRNA accumulation at about 8 h on average. IL-2R mRNA expression was an exception, showing a prolonged increase (72 h). The general profiles of expression of the five cytokine genes were similar but not identical, suggesting some shared regulatory mechanisms. When responses to four additional stimuli (pokeweed mitogen, Candida albicans, and IL-2 at high and low doses) were compared, similar profiles of cytokine gene expression were found. Thus, the various stimuli caused induction of all cytokines with quantitative, not qualitative, differences. Altogether, the present data are useful for defining the kinetics of gene expression for key cytokines in response to standard immune-cell stimuli.

Monocytes and dendritic cells are the primary sources of interleukin 37 in human immune cells

“…**P , 0.01 for vehicle + BFA compared with anti-CD3 + BFA. and IL6 and the anti-inflammatory IL10 by anti-CD3-stimulated PBMCs [37] further support the hypothesis that the delayed IL1F7 mRNA expression observed here originated from cells other than T cells.…”

Dynamics of cytokine gene expression in peripheral blood mononuclear cells of indigenous and exotic breeds of pigs in India

“…We also observed that even unstimulated PBMCs expressed low levels of all cytokine transcripts studied. Our results are consistent with observations for IFN-c expression in unstimulated spleen cells (Iizawa et al 1992), IL-2 and IFN-c expression in rat resting T-cell clones (Zhao et al 1994), and IL-2, IFN-c, IL-6, IL-10 and TNF-a transcripts in unstimulated human PBMCs (Fan et al 1998). Beta actin expression was little affected by mitogenic stimulation of PBMCs at any point of time examined in both Zovawk and LWY pigs, thus serving as the reference gene (Facci et al 2011) in our real-time PCR assay.…”

“…Upon PBMCs activation, coordinated secretion of various cytokines control proliferation, differentiation, and function of cells and are crucial for regulation of immune response (Corry et al 1996;Fan et al 1998;Dar et al 2015). Our results depict significant increase in all the cytokines genes' expression assayed except IL-2, within 1 to 3 h of PHA-P stimulation.…”

“…As a mixed population of T-lymphocytes, B-lymphocytes, natural killer cells, monocytes, macrophages and dendritic cells, PBMCs represent key cells involved in the genetic expression of various cytokines leading to innate and adaptive immune and inflammatory responses. Among mitogens, phytohemagglutinin (PHA) is a mitogen that predominantly stimulates T-lymphocytes within the PBMC population (Mire-Sluis et al 1987;Fan et al 1998;Abdalla et al 2003). In addition, the analysis of cytokine profiles is of fundamental importance for a better understanding of the functional properties of immune cells in physiological and pathological conditions.…”

Long‐term effects of idiotype vaccination on the specific T‐cell response in peripheral blood and bone marrow of multiple myeloma patients

“…Id‐specific mRNA gene expression was expressed as a ratio between the relative increase of a gene induced by the autologous Id compared with that of the isotype control (fold increase ratio). The expression of each gene was calculated in relation to unactivated cells after normalization against β‐actin (28, 29). An Id‐specific response was considered when the relative fold increase of a gene induced by the autologous Id was higher than the isotypic control.…”